Nuclei-specific differences in nerve terminal distribution, morphology, and development in mouse visual thalamus
- Sarah Hammer†1, 2,
- Gabriela L Carrillo†1, 3,
- Gubbi Govindaiah†6,
- Aboozar Monavarfeshani1, 4,
- Joseph S Bircher1, 5,
- Jianmin Su1,
- William Guido6 and
- Michael A Fox1, 4, 5Email author
© Hammer et al.; licensee BioMed Central Ltd. 2014
Received: 24 April 2014
Accepted: 25 June 2014
Published: 10 July 2014
Mouse visual thalamus has emerged as a powerful model for understanding the mechanisms underlying neural circuit formation and function. Three distinct nuclei within mouse thalamus receive retinal input, the dorsal lateral geniculate nucleus (dLGN), the ventral lateral geniculate nucleus (vLGN), and the intergeniculate nucleus (IGL). However, in each of these nuclei, retinal inputs are vastly outnumbered by nonretinal inputs that arise from cortical and subcortical sources. Although retinal and nonretinal terminals associated within dLGN circuitry have been well characterized, we know little about nerve terminal organization, distribution and development in other nuclei of mouse visual thalamus.
Immunolabeling specific subsets of synapses with antibodies against vesicle-associated neurotransmitter transporters or neurotransmitter synthesizing enzymes revealed significant differences in the composition, distribution and morphology of nonretinal terminals in dLGN, vLGN and IGL. For example, inhibitory terminals are more densely packed in vLGN, and cortical terminals are more densely distributed in dLGN. Overall, synaptic terminal density appears least dense in IGL. Similar nuclei-specific differences were observed for retinal terminals using immunolabeling, genetic labeling, axonal tracing and serial block face scanning electron microscopy: retinal terminals are smaller, less morphologically complex, and more densely distributed in vLGN than in dLGN. Since glutamatergic terminal size often correlates with synaptic function, we used in vitro whole cell recordings and optic tract stimulation in acutely prepared thalamic slices to reveal that excitatory postsynaptic currents (EPSCs) are considerably smaller in vLGN and show distinct responses following paired stimuli. Finally, anterograde labeling of retinal terminals throughout early postnatal development revealed that anatomical differences in retinal nerve terminal structure are not observable as synapses initially formed, but rather developed as retinogeniculate circuits mature.
Taken together, these results reveal nuclei-specific differences in nerve terminal composition, distribution, and morphology in mouse visual thalamus. These results raise intriguing questions about the different functions of these nuclei in processing light-derived information, as well as differences in the mechanisms that underlie their unique, nuclei-specific development.
KeywordsRetina Thalamus Retinogeniculate Lateral geniculate nucleus Axon Retinal terminal Nerve terminal
The visual thalamus of rodents has served as an important model for exploring the cellular and molecular mechanisms that underlie neural circuit formation. The overwhelming majority of these studies have focused on inputs to and projections from the dorsal lateral geniculate nucleus (dLGN). Relay neurons within dLGN receive strong glutamatergic inputs from retinal ganglion cells (RGCs) and serve as the principle conduit of visual signaling to the cortex. However, relay neurons do not act as passive relays of visual information. The gain of retinogeniculate signal transmission is modulated by nonretinal inputs to dLGN. These nonretinal inputs arise from visual cortex, pretectum, brainstem, thalamic reticular nuclei, and local dLGN interneurons, and they far outnumber the more powerful retinal inputs [1, 2]. In fact, nonretinal inputs account for as much as 95% of the nerve terminals in dLGN [1, 3–6].
Differences in the functional properties of inputs to dLGN translate into distinct neurochemical and ultrastructural differences in retinal and nonretinal synapses in dLGN. Retinal, cortical, brainstem and inhibitory nerve terminals in dLGN all contain distinct neurotransmitter synthesizing enzymes and synaptic vesicle associated transporter proteins necessary for their specific functions [4, 7–12]. At the ultrastructural level, several types of nerve terminals have been described in dLGN, defined by anatomical features such as terminal morphology, synaptic vesicle shape, and mitochondrial appearance [6, 13–16]. One set of nerve terminals contain flattened, oval shaped synaptic vesicles, a hallmark feature of inhibitory GABAergic terminals, and are therefore called F terminals [2, 14, 17]. In dLGN, GABAergic terminals arise from the thalamic reticular nucleus and local inhibitory interneurons [7, 18, 19]. Other classes of terminals in dLGN contain round (or rather spherical) synaptic vesicles, and these represent excitatory and modulatory inputs from the retina, cortex, brainstem, pretectum and superior colliculus . Glutamatergic retinal terminals are distinguished from all other round-vesicle containing terminals based on their exceptionally large size and pale-colored mitochondria [6, 13, 14, 20]. For this reason retinal terminals in dLGN are termed RLP terminals (for r ound synaptic vesicles, l arge terminal size, and p ale-colored mitochondria). In contrast, nonretinal excitatory/modulatory nerve terminals, which far outnumber RLPs, are termed RSD terminals for their round synaptic vesicles, small terminal size, and dark-colored mitochondria [3, 4, 6]. In addition to terminal size and mitochondrial appearance, retinal terminals in dLGN are distinguishable from other terminal types as they exhibit complex synaptic arrangements with F terminals, they cluster into complex synaptic zones encapsulated by glial processes (and arrangement termed a glomeruli), and they typically contain invaginations of spine-like structures from either dendrites of dLGN principle neurons or F terminals [2, 14, 20].
While synaptic organization and morphology in dLGN have been thoroughly explored, other retino-recipient regions of mouse thalamus have received far less attention. Adjacent to the dLGN are the ventral lateral geniculate nucleus (vLGN) and the intergeniculate leaflet (IGL), two thalamic nuclei that receive and process light derived information from the retina. It is important to note that while retinal axons innervate all regions of dLGN and IGL, only the external division of the rodent vLGN receives retinal afferent . For simplicity sake we will be referring to this external division of vLGN throughout this manuscript. In contrast to dLGN, which relays image-forming visual information to the primary visual cortex, vLGN and IGL contribute to functions of the non-image forming accessory visual system, such as irradiance detection, visuomotor responses, and circadian photo-entrainment [22, 23]. Source of inputs to vLGN and IGL are similar to that of dLGN, which includes inputs from retina, cortex, superior colliculus, thalamic reticular nucleus, and local interneurons [22, 24]. However, it is becoming increasingly clear that different classes of neurons from these nuclei project to distinct nuclei of visual thalamus. For example, while classes of image-forming RGCs, which transmit information regarding contrast, color and motion, project to dLGN, classes of non-image forming RGCs project to vLGN and IGL [23, 25–34]. Likewise, distinct classes of cortical neurons innervate dLGN and vLGN. Layer VI cortical neurons project to dLGN, whereas layer V cortical neurons project to vLGN [24, 35, 36]. In addition to differences in their inputs, projections from relay neurons in dLGN and vLGN widely differ. In contrast to the thalamocortical projections originating from dLGN, projections from vLGN do not enter cortex and instead innervate an array of ipsi- and contralateral structures within hypothalamus, thalamus and midbrain .
Despite this knowledge of the circuits associated with the more ventrally located nuclei of rodent visual thalamus, we lack important information regarding the organization, distribution and morphology of nerve terminals in this region. We therefore explored and compared the distribution, morphology and physiology of nerve terminals in vLGN and dLGN using immunolabeling, mouse genetics, anterograde axonal labeling, serial block face scanning electron microscope (SBFSEM) and whole-cell patch recordings. Our results show that nerve terminal composition not only differs between visual thalamic nuclei, but that, retinal terminal development, morphology and physiology also differ significantly in these nuclei.
Results and discussion
Distribution of inhibitory and modulatory terminals in mouse visual thalamic nuclei
A more striking difference in the distribution of inhibitory nerve terminals was apparent following GAD65-immunostaining. GAD65 was robustly present in vLGN and IGL but was lowly expressed in dLGN (Figure 1E,F; see ). In many brain regions, GAD65 and GAD67 are present in different subsets of inhibitory nerve terminals [38–40]. This appears to be the case in both dLGN and IGL since GAD67+/GAD65- terminals were present in dLGN and GAD67-/GAD65+ terminals were present in IGL (Figure 1A,C-F and ). Since terminals in vLGN were labeled with antibodies against both GAD67 and GAD65, it remains unclear whether these terminals co-express both enzymes, or whether these represent two unique populations of inhibitory nerve terminals in vLGN.To confirm differences in inhibitory terminal distribution in dLGN and vLGN, we labeled inhibitory terminals with antibodies against VGAT. These results closely resembled those obtained with GAD67 immunolabeling. VGAT-positive inhibitory terminal size appeared similar in dLGN and vLGN, but the density of these terminals was higher in vLGN (Figure 1G,H).
We next explored the distribution of cholinergic nerve terminals in vLGN and dLGN [4, 41]. These modulatory inputs arise from several brainstem nuclei. We immunolabeled these terminals with antibodies directed against Vesicle Associated Choline Transporter (VAChT), which is required to load synaptic vesicles with acetylcholine. VAChT-positive terminals appeared similar in size in vLGN and dLGN (Figure 1B,I,J), and the density of cholinergic terminals in vLGN and dLGN was similar (in vLGN 4.3% ± 0.9% of images were occupied with VAChT-IR versus 5.3% ± 0.5% in dLGN. Data are shown as mean ± SEM. P = 0.14 by Student’s t-test). Thus, while significant differences were noted in the composition and density of inhibitory terminals in mouse visual thalamic nuclei, the size and density of cholinergic inputs appears to lack nuclei-specific differences.
Distribution of excitatory nerve terminals in mouse visual thalamic nuclei
In contrast to the dense cortical inputs in dLGN, VGluT1-positive terminals sparsely populated vLGN (Figure 2A,D). To test whether VGluT1-containing cortical terminals in dLGN and vLGN originated from distinct cortical layers, we examined cortical projections in Golli-tau-gfp transgenic mice, in which layer VI cortical neurons (but not layer V neurons) are labeled with Green Fluorescent Protein (GFP) [35, 36, 42]. As was the case for VGluT1-IR, tau-GFP distribution was so dense in adult dLGN that individual nerve terminals could not be distinguished, even at high magnification in single optical sections of confocal images (Figure 2E,G). In fact, the only regions of dLGN in Golli-tau-gfp transgenic mice that appeared devoid of GFP (in single optical sections) were regions containing cell somas, VGluT2-IR retinal terminals , or blood vessels (Figure 2G,H and JS and MAF, unpublished observations). In contrast, tau-GFP-positive projections were absent from vLGN (Figure 2F). This suggested that VGluT1-positive terminals in mouse vLGN do not arise from cortical layer VI. Alternative possibilities were that VGluT1-positive terminals in vLGN arose from cortical layer V [24, 43], from retinal projections, or from the superior colliculus, a third source of glutamatergic inputs to visual thalamus . We ruled out the possibility that VGluT1-positive terminals arose from retinal projections since they persisted in vLGN of Math5 -/- mutant mice, which lack retinal inputs to the thalamus (JS and MAF, unpublished observations) [36, 44, 45]. It was also unlikely that VGluT1-positive terminals in vLGN arose from superior colliculus since these neurons express VGluT2 and not VGluT1 . Taken together with studies in rat thalamus, these data lead us to speculate that VGluT1-positive terminals in mouse vLGN originate from layer V cortical neurons [24, 43]. In contrast to modulatory inputs that originate from cortical layer VI, inputs from layer V generate strong, driver-like input to higher order thalamic nuclei [1, 46, 47]. This raises the possibility that VGluT1-positive corticogeniculate synapses provides a primary excitatory drive for vLGN neurons.
Like VGluT1-IR, the distribution of VGluT2-IR differed significantly in distinct nuclei of the mouse visual thalamus. Little immunoreactivity was observed in IGL or in the non-retino-recipient regions of vLGN (i.e. the internal division of the vLGN) . Robust VGluT2-IR was detected in both vLGN and dLGN, however the patterns of immunoreactivity in these nuclei differed. As noted previously, the size of VGluT2-containing terminals varied between these regions, with larger terminals predominating in dLGN (Figure 2I-L) . Not only were dLGN VGluT2-positive terminals larger than their vLGN counterparts, but they appeared larger than all other types of terminals analyzed here (for example, in dLGN VGluT2-positive terminals averaged 2.43 μm2 ± 0.38 μm2 (SD) whereas VAChT-positive terminals averaged 0.70 μm2 ± 0.06 μm2 and GAD67-positive terminals averaged 1.21 μm2 ± 0.11 μm2; see Figure 2K,L). Interestingly, despite terminals being dramatically larger in dLGN, a significantly larger fraction of each confocal image was occupied by VGluT2-IR in vLGN (15.6% ± 0.5% of vLGN images were occupied by VGluT2-IR versus 8.3% ± 0.3% in dLGN; P <0.0001 by Student’s t-test).
Based on previous reports showing VGluT2 expression by RGCs , we interpreted these findings to indicate that retinal terminals are smaller but more densely distributed in vLGN. To ensure that VGluT2-containing terminals originated from the retina, we assessed VGluT2-IR in Math5 -/- mutant mice. Few VGluT2-positive terminals were observed in Math5 -/- mutant vLGN and dLGN. We interpret this data to indicate that: 1). The vast majority of VGluT2-positive terminals in mouse visual thalamus were retinal terminals; 2). A small cohort of nonretinal, VGluT2-positive terminals existed in both vLGN and dLGN. These terminals, which we suspect arise from superior colliculus (based on the robust expression of VGluT2 by collicular projection neurons) , appeared similar in size in both vLGN and dLGN and are smaller than dLGN retinal terminals.
Nuclei-specific differences in retinal terminal morphology in mouse visual thalamus
We interpret differences in VGluT2-IR in vLGN and dLGN to reflect differences in retinal nerve terminal morphology in these regions. However, since VGluT2-IR labeled synaptic vesicle pools and not entire nerve terminals, an alternative possibility was that the quantity or distribution of synaptic vesicles (and not terminal size) differed in vLGN and dLGN. To distinguish between these possibilities we labeled retinal axons and their terminals with two orthologous techniques: transgenic expression of fluorescent reporter proteins and anterograde labeling by intraocular injection of fluorescently conjugated tracer molecules.
Retino-recipient nuclei in a wide variety of mammalian brains are subdivided into discrete laminae (or regions), a feature that facilitates the flow of visual information through distinct parallel pathways in the brain. Although laminae are not readily identifiable by cyto-architectural analysis, it is becoming increasingly clear that retinal terminals are targeted into ‘hidden’ laminae of the rodent dLGN ([50–52]) In mice, these ‘hidden’ subdivisions occur along the lateral-medial axis and are referred to as the ‘shell’ and ‘core’ of dLGN . As we analyzed retinal terminal size throughout LGN, we noted a small, lateral ‘shell’-like region of vLGN that contained retinal terminals that were morphologically distinct from the rest of vLGN and IGL (Figure 5A-G). This morphologically distinct region of vLGN was present only in caudal sections of vLGN; therefore, we refer to it as the lateral shell of caudal vLGN (lcvLGN). CTB-labeled terminals in lcvLGN were significantly larger than those in IGL, in more medial ‘core’-like regions of caudal vLGN, and in both medial and lateral regions of rostral vLGN (Figure 5H). An interesting feature of terminals in lcvLGN that we failed to detect in any other region of the visual thalamus (or other retino-recipient nuclei) was that retinal terminal size correlated with eye of origin. Terminals in the lcvLGN that originated from the contralateral retina appeared consistently larger than those in other regions of vLGN of IGL (Figure 5E-G). In contrast, terminals in the lcvLGN that originated from the ipsilateral retina appeared small and consistent in size to the more typical retinal terminals in IGL and vLGN terminals (Figure 5E-G). At least two classes of direction-selective RGCs have been identified that not only project to both dLGN and lateral regions of caudal vLGN, but exclusively project to contralateral thalamus [27, 53]. It is possible that axons from these classes of direction-selective RGCs generate these large terminals in lcvLGN.
Ultrastructural analysis of retinal terminals in mouse visual thalamus
Retinal terminals are clustered into dense ‘synaptic islands’ (i.e. glomeruli) in dLGN. Whether such complex arrangements exist in mouse vLGN remains unresolved (but see ). Therefore, one possibility is that dLGN retinal terminals appeared larger following immunohistochemistry (IHC), genetic labeling or anterograde labeling due to the inability of confocal microscopy to fully resolve individual terminals in these complex synaptic arrangements. To address this possibility, we examined retinal terminal morphology with serial block face scanning electron microscopy (SBFSEM), a technique that permits the tracing of ultrastructural morphology of nerve terminals through relatively large volumes of serially sectioned tissue. Four samples of vLGN and dLGN from adult wild-type mice were analyzed by SBFSEM. Each sample set represented a 40 μm by 40 μm by 15 μm volume of tissue.
We used identical criteria to characterize the ultrastructural morphology of retinal terminals in vLGN. Similar to dLGN, pale mitochondria-containing terminals made multiple synaptic contacts and were closely associated with inhibitory terminals (Figure 7J and SH, JSB, and MAF, unpublished observation). Pale mitochondria-containing retinal terminals in vLGN also existed in isolation or in clusters around dendrites, although these clusters were often not encapsulated by glial cells as was the case in dLGN (Figure 7C,D). However, despite these similarities, retinal terminals appeared strikingly different in vLGN. First, they appeared significantly smaller in vLGN than dLGN (Figures 7 and 8). After tracing each pale-mitochondria containing terminal in its entirety, we measured the diameter of the widest portion of each presynaptic bouton. The mean width of retinal terminals in dLGN was 2.88 μm ± 0.11 μm versus 1.55 μm ± 0.04 μm in vLGN (P <0.00001 by Student’s t-test. n = 148 boutons in vLGN; n = 111 boutons in dLGN. Data are mean ± SEM) (Figure 7E). Second, retinal terminals in vLGN appeared less morphological complex, as <10% of these terminals contained finger-like invaginations from postsynaptic target cells (Figure 7F,J-N). These results are in line with previous studies in cats, which revealed smaller retinal terminals and a lack of typical, glial encapsulated glomeruli in vLGN .
In serially tracing entire retinal arbors with SBFSEM datasets, we observed one addition difference in presynaptic axons and boutons in these nuclei. Axonal arbors traced in vLGN (n = 44) contained on average more terminal boutons than axons traced in dLGN (Figure 7). Figure 7M shows a distribution plot of the numbers of terminal boutons in each axon traced in vLGN and dLGN. Axons in dLGN contained approximately 40% fewer terminal boutons than those traced in vLGN (1.85 ± 0.12 boutons per axon in dLGN (n = 59 axons) versus 3.29 boutons per axon in vLGN (n = 44). Data are mean ± SEM. P <0.00005 by Student’s t-test).
Distinct patterns of retinogeniculate transmission in ventral lateral geniculate nucleus and dorsal lateral geniculate nucleus
At glutamatergic synapses, a structure-function relationship has been described in which nerve terminal size significantly influences the functional strength of information transfer . Anatomically large glutamatergic synapses with little convergence on postsynaptic target cells (for example, dLGN RLPs) produce large amplitude excitatory postsynaptic currents (EPSCs), while smaller terminals elicit weaker postsynaptic responses. Based on results described above, we postulated that the synaptic strength of retinogeniculate synapses should vary between mouse vLGN and dLGN.
By contrast, responses in vLGN evoked by optic tract stimulation were fivefold smaller (Figure 9A). Moreover, EPSCs showed a graded increase in amplitude with a progressive increase in stimulus intensity (Figure 9B), indicating a higher degree of retinal convergence on vLGN neurons . This result may, at least in part, explain the increased density of retinal terminals observed following VGluT2-IHC and CTB labeling in vLGN (Figures 2, 4 and 5) and the increased number of boutons per retinal axon observed with SBFSEM in vLGN (Figure 8). In contrast to dLGN, retinogeniculate synapses in vLGN exhibit much less depression following repetitive optic tract stimulation (Figure 9C-E).
Taken together, anatomical and physiological comparisons between dLGN and vLGN suggest that retinal terminals in vLGN do not seem to possess the hallmark features of a driver-like synapse. Instead, they seem to have features consistent with modulatory or Class 2-type synapses. Retinal terminals in vLGN appear smaller in size than those in dLGN, produce weak EPSCs, and exhibit higher levels of convergence on vLGN neurons. Moreover, studies in rodents further suggest that retinal terminals contact distal regions of vLGN neuron dendrites  whereas those in dLGN reside just proximal to somata . While all of these features more closely resemble features associated with modulatory glutamatergic inputs rather than driver synapses, it is important to note that vLGN synapses responses did not show paired pulse facilitation, a feature common to most modulatory synapses (Figure 9E). Thus, perhaps retinal synapses in vLGN represent a hybrid between these two terminal types.
Development of retinal terminal morphology in ventral lateral geniculate nucleus and dorsal lateral geniculate nucleus
At present it remains unclear whether differences in terminal development in vLGN and dLGN are intrinsic characteristics of the classes of neurons that project to either nuclei or whether target-derived cues differentially transform terminals in nuclei-specific fashions. A case could be made that terminal morphology is an intrinsic feature of axons if the larger retinal terminals observed in lcvLGN do in fact originate from the same direction selective RGCs that project to lateral regions of dLGN [27, 53]. However, both single axon tracing studies and observations of genetically labeled RGCs indicate that dLGN-projecting retinal axons branch to innervate other retino-recipient nuclei that lack anatomically large retinal terminals [33, 53, 56]. This suggests that identity alone does not predetermine terminal size.
An elegant series of experiments by Frost and colleagues supports the notion of target-derived cues capable of differentially shaping retinal terminal maturation and morphology in different thalamic nuclei [62–65]. In these studies, the misrouting of retinal axons into non-retino-recipient regions of dorsal thalamus resulted in retinal terminals adopting nonretinal-like morphologies [64, 65]. In fact, data from our own studies in mice lacking canonical Reelin receptors  support the notion that local environmental cues released by nuclei-specific cells pattern the targeting and differentiation of nerve terminals in mouse visual thalamus. At present, the nature of such cues remains unclear.
In the present study, we examined the composition, distribution and morphology of nerve terminals in three adjacent, retino-recipient nuclei in the mouse thalamus. Results demonstrate that despite all receiving, processing and relaying light-derived signals from the retina, each nucleus contains distinct sets of neurochemically-defined nerve terminals. Inhibitory terminals (which contained GAD67, GAD65, or VGAT) are most abundant in vLGN. Cortical inputs are most abundant in dLGN and appear to originate from a different layer of cerebral cortex in each visual thalamic nucleus. And most surprising was our discovery that retinal terminals vary significantly in their abundance and morphology in these nuclei. While retinal terminals appear significantly larger and more complex in dLGN, they appear more abundant in vLGN. Differences in retinal terminals are not limited to their anatomy, since retinogeniculate synaptic responses varied significantly in vLGN and dLGN. While retinogeniculate synapses in dLGN exhibited all of the hallmark features of driver inputs, retinal synapses in vLGN elicited weaker postsynaptic responses and displayed features associated with higher levels of convergence on target neurons. These results raise intriguing questions about the different functions of these nuclei in processing light-derived information, as well as differences in the mechanisms that underlie their unique, nuclei-specific development.
Wild-type C57 mice were obtained from Charles River (Wilmington, MA). Math5-cre and Rosa:lox-stop-lox:tdt (Jackson Laboratory Stock #007905; referred to here as Rosa-stop-tdt) mice were generously provided Dr. C.K. Chen (Baylor College of Medicine, Houston TX). All analyses conformed to National Institutes of Health guidelines and protocols approved by the Virginia Polytechnic Institute and State University Institutional Animal Care and Use Committees.
Antibodies used in these studies
Description of immunogen
Dilution for immunohistochemistry
Glutamate Decarboxylase 67 (Gad 67)
Glutamate Decarboxylase 65 (Gad 65)
Synthetic peptide containing amino acids 572 to 585 of rat Gad65
Vesicular Glutamate Transporter 1 (VGluT1)
Recombinant fusion protein consisting of amino acids 456 to 560 of rat VGlut2
Synaptic Systems #135303
Vesicular Glutamate Transporter 2 (VGluT2)
Recombinant fusion protein consisting of amino acids 510 to 582 of rat VGlut2
Synaptic systems #135403
Vesicular GABA Transporter (VGAT)
Synthetic peptide containing amino acids 510 to 525 of rat GABA
Synaptic systems #131103
Vesicular Acetyl-choline Transporter (VAChT)
Guinea Pig IgG
Fusion protein consisting of amino acids 475 to 530 of rat VAChT
Synaptic systems #139105
Immunohistochemistry (IHC) was performed on 16-μm coronal cryosectioned tissues as described previously . Briefly, tissue slides were allowed to air dry for 15 minutes before being incubated with blocking buffer (2.5% normal goat serum, 2.5% bovine serum albumin, 0.1% Triton X-100 in PBS) for 30 minutes. Primary antibodies were diluted in blocking buffer and incubated on tissue sections for overnight at 4°C. On the following day, tissue slides were washed in PBS and secondary antibodies diluted 1:1000 in blocking buffer were applied to slides for 1 hour at room temperature. After thoroughly washing in PBS, tissue slides were coverslipped with VectaShield (Vector Laboratories, Burlingame, CA, USA). Images were acquired on a Zeiss LSM 700 confocal microscope. When comparing different ages of tissues or between genotypes, images were acquired with identical parameters. A minimum of three animals (per age) was compared in all IHC experiments. Image analysis was performed in ImageJ using identical parameters for all image sets. Briefly, single color IHC images were binarized and inverted with the ‘Threshold’ tool with NIH ImageJ (Image > Adjust > Threshold tool) (http://imagej.nih.gov/ij/). Threshold values for each set of immunofluorescent images (i.e. for each antibody applied) were selected based on the histogram analysis of dLGN images and were then applied to images from both dLGN and vLGN. Threshold values were as follows: VGluT2-IR - 30; GAD67-IR - 45; VAChT-IR - 30. Terminal size analysis was determined with the ‘Analyze Particle’ feature in ImageJ (Analyze > Analyze Particles tool) and the fraction of images containing immunoreactivity was determined with the ‘Measure’ feature (Analyze > Measure tool). Image analysis was performed on three to five randomly distributed regions of dLGN and vLGN from each hemisphere of at least three wild-type P60 mice. Statistical analysis of quantified images was performed in StatPlus (Analyst Software, Inc.; http://www.analystsoft.com/en/).
Intraocular injections of anterograde tracers
Intraocular injection of cholera toxin subunit B (CTB) conjugated to Alexa Fluor 488 or Alexa Fluor 594 (Invitrogen/Life Technologies, Grand Island, NY) was performed as described previously [37, 49]. Briefly, mice were anesthetized with hypothermia (<P7) or by isoflurane vapors (>P7). The sclera was pierced with a sharp-tipped glass pipette and excess vitreous was drained. Another pipette, filled with a 0.1 to 0.2% solution of CTB, was inserted into the hole made by the first pipette. The pipette containing the CTB was attached to a Picospritzer and a prescribed volume (1 to 3 μl at P3 and 3 to 5 μl for ages > P10) of solution was injected into the eye. After 2 d, mice were euthanized, transcardially perfused with PBS and 4% paraformaldehyde, and brains were post-fixed in 4% paraformaldehyde for 12 hours. Fixed brains were coronally sectioned (80 to 100 μm) on a vibratome (Microm HM 650 V, Thermo Scientific) and mounted in ProLong Gold (Invitrogen/Life Technologies, Grand Island, NY). Retinal projections were analyzed from between three to six animals for each age. Images were acquired on a Zeiss LSM 700 confocal microscope. Terminal size was measured manually in Image J in single optical sections of confocal images from three to five randomly distributed regions of dLGN, IGL, lcvLGN and vLGN from each hemisphere of at least three wild-type mice (per age). Statistical analysis of quantified images was performed in StatPlus (Analyst Software, Inc.; http://www.analystsoft.com/en/).
Serial block face scanning electron microscopy
Mice were transcardially perfused sequentially with PBS and 4% paraformaldehyde/2% glutaradehyde in 0.1 M cacodylate buffer. Brains were immediately removed, vibratomed (300 μm coronal sections) and vLGN and dLGN were dissected. Tissues were then stained, embedded, sectioned and imaged by Renovo Neural Inc. (Cleveland, OH). Images were acquired at a resolution of 5 nm/pixel and image sets included >200 serial sections (with each section representing 75 nm in the z axis). SBFSEM data sets were 40 μm × 40 μm × approximately 15 μm. Four data sets were analyzed for each region (from a total of three wild-type mice). Data sets were analyzed in TrakEM2 (http://fiji.sc/TrakEM2) . Retinal terminals were identified by the presence of synaptic vesicles and pale mitochondria. Large terminal size was not used as an identifying criteria for retinal terminals. Analysis of data sets was performed independently by two researchers, blind to the tissue of origin for each data set, to ensure unbiased results. Additionally, each analyzed separate sample sets of dLGN and vLGN to ensure terminals were not double counted. Statistical analysis of quantified images was performed in StatPlus (Analyst Software, Inc.; http://www.analystsoft.com/en/).
Whole-cell patch recordings in visual thalamus
Whole-cell patch recordings were performed as previously described with modifications . P35 wild-type mice were anesthetized with isoflurane, decapitated and brains were rapidly immersed in an ice-cold, oxygenated (95% O2/5% CO2) solution containing the following (in mM): 26 NaHCO3, 234 sucrose, 10 MgCl2,11 glucose, 2.5 KCl, 1.25 NaH2 PO4, 2 CaCl2. Coronal sections (300 μm) containing vLGN and dLGN were cut on a vibratome and were incubated in artificial cerebral spinal fluid (ACSF; in mM: 126 NaCl, 2.5 KCl, 1.25 NaH2PO4, 2.0 MgCl2, 26 NaHCO3, 2 CaCl2, 2 MgCl2 and 10 glucose, saturated with 95% O2/5% CO2, pH 7.3) at 32°C for 25 min and then room temperature. Individual slices were transferred to a recording chamber maintained at 32°C and perfused continuously at a rate of 2.5 ml/min with oxygenated artificial cerebrospinal fluid (ACSF). Patch electrodes were pulled using a two-step puller (Narishige) from borosilicate glass and filled with a solution containing the following (in mM): 117 K-gluconate, 13 KCl,1 MgCl2, 0.07 CaCl2, 0.01 EGTA, 10 HEPES, 2 Na-ATP, and 0.4 Na-GTP (pH 7.3, 290 osmol/L). The final tip resistance of filled electrodes was 3 to 6 MΩ.
Synaptic responses were evoked by electrical stimulation of the optic tract (OT) using bipolar tungsten electrodes (0.5 MΩ; A-M Systems, Carlsborg, WA) positioned at the caudal ventral border of the LGN [79, 80]. Whole-cell recordings were done in voltage-clamp mode at a holding potential of -70 mV and in the presence of SR95531 (10 μM) and the GABAB receptor antagonist 3-minopropyl diethoxymethyl phosphinic acid (10 μM; Tocris Bioscience, Bristol, UK). Single synaptic responses were evoked every 20 s across a range of stimulus intensities (25 to 125 mA). Trains of paired stimuli were delivered (using the stimulus that produced the maximal response) at 20 Hz (50 ms interpulse interval). Whole-cell recordings were obtained using Multiclamp 700 B amplifier (Molecular Devices, Sunnyvale, CA). Data were filtered at 2.5 KHz, digitized at 10 kHz using an interface unit (Digidata 1440A, Molecular Devices), and stored on a computer. Current traces were filtered at 5 kHz; events were detected and amplitudes measured using pClamp 10 software (Molecular Devices).
artificial cerebrospinal fluid
cholera toxin subunit B
dorsal lateral geniculate nucleus
excitatory postsynaptic currents
Glutamate decarboxylase 65
Glutamate decarboxylase 67
green fluorescent protein
intergeniculate nucleus (leaflet)
lateral shell of caudal ventral lateral geniculate nucleus
lateral geniculate nucleus
olivary pretectal nucleus
retinal ganglion cells
round synaptic vesicles, large terminal size, and pale-colored mitochondria
round synaptic vesicles, small terminal size, and dark-colored mitochondria
serial block face scanning electron microscope
the suprachiasmatic nucleus
Vesicular Acetyl-choline Transporter
Vesicular GABA Transporter
Vesicular Glutamate Transporter 1
Vesicular Glutamate Transporter 2
ventral lateral geniculate nucleus.
This work was supported by the National Institutes of Health (NIH) - National Eye Institute Grants EY021222 (MAF) and EY012716 (WG). We thank Dr. C.K. Chen (Baylor College of Medicine) for providing math5-cre and rosa-stop-tdt mice, Dr. A.T.Campagnoni (UCLA) for providing golli-tau-gfp mice, and Dr. S.W. Wang (University of Texas Health Science Center) for providing math5 -/- mice.
- Sherman SM, Guillery RW: The role of the thalamus in the flow of information to the cortex. Philos Trans R Soc Lond B Biol Sci. 2002, 357: 1695-1708.PubMed CentralView ArticlePubMedGoogle Scholar
- Sherman SM: Interneurons and triadic circuitry of the thalamus. Trends Neurosci. 2004, 27: 670-675.View ArticlePubMedGoogle Scholar
- Erisir A, Van Horn SC, Bickford ME, Sherman SM: Immunocytochemistry and distribution of parabrachial terminals in the lateral geniculate nucleus of the cat: a comparison with corticogeniculate terminals. J Comp Neurol. 1997, 377: 535-549.View ArticlePubMedGoogle Scholar
- Erisir A, Van Horn SC, Sherman SM: Relative numbers of cortical and brainstem inputs to the lateral geniculate nucleus. Proc Natl Acad Sci U S A. 1997, 94: 1517-1520.PubMed CentralView ArticlePubMedGoogle Scholar
- Van Horn SC, Erisir A, Sherman SM: Relative distribution of synapses in the A-laminae of the lateral geniculate nucleus of the cat. J Comp Neurol. 2000, 416: 509-520.View ArticlePubMedGoogle Scholar
- Bickford ME, Slusarczyk A, Dilger EK, Krahe TE, Kucuk C, Guido W: Synaptic development of the mouse dorsal lateral geniculate nucleus. J Comp Neurol. 2010, 518: 622-635.PubMed CentralView ArticlePubMedGoogle Scholar
- Fitzpatrick D, Penny GR, Schmechel DE: Glutamic acid decarboxylase-immunoreactive neurons and terminals in the lateral geniculate nucleus of the cat. J Neurosci. 1984, 4: 1809-1829.PubMedGoogle Scholar
- Montero VM, Singer W: Ultrastructural identification of somata and neural processes immunoreactive to antibodies against glutamic acid decarboxylase (GAD) in the dorsal lateral geniculate nucleus of the cat. Exp Brain Res. 1985, 59: 151-165.View ArticlePubMedGoogle Scholar
- Gabbott PL, Somogyi J, Stewart MG, Hamori J: GABA-immunoreactive neurons in the dorsal lateral geniculate nucleus of the rat: characterization by combined Golgi-impregnation and immunocytochemistry. Exp Brain Res. 1986, 61: 311-322.PubMedGoogle Scholar
- Moore RY, Speh JC: GABA is the principal neurotransmitter of the circadian system. Neurosci Lett. 1993, 150: 112-116.View ArticlePubMedGoogle Scholar
- Fremeau RT, Troyer MD, Pahner I, Nygaard GO, Tran CH, Reimer RJ, Bellocchio EE, Fortin D, Storm-Mathisen J, Edwards RH: The expression of vesicular glutamate transporters defines two classes of excitatory synapse. Neuron. 2001, 31: 247-260.View ArticlePubMedGoogle Scholar
- Land PW, Kyonka E, Shamalla-Hannah L: Vesicular glutamate transporters in the lateral geniculate nucleus: expression of VGLUT2 by retinal terminals. Brain Res. 2004, 996: 251-254.View ArticlePubMedGoogle Scholar
- Guillery RW: The organization of synaptic interconnections in the laminae of the dorsal lateral geniculate nucleus of the cat. Z Zellforsch Mikrosk Anat. 1969, 96: 1-38.View ArticlePubMedGoogle Scholar
- Lund RD, Cunningham TJ: Aspects of synaptic and laminar organization of the mammalian lateral geniculate body. Invest Ophthalmol. 1972, 11: 291-302.PubMedGoogle Scholar
- Robson JA, Mason CA: The synaptic organization of terminals traced from individual labeled retino-geniculate axons in the cat. Neuroscience. 1979, 4: 99-111.View ArticlePubMedGoogle Scholar
- Hamos JE, Van Horn SC, Raczkowski D, Sherman SM: Synaptic circuits involving an individual retinogeniculate axon in the cat. J Comp Neurol. 1987, 259: 165-192.View ArticlePubMedGoogle Scholar
- Famiglietti EV: Dendro-dendritic synapses in the lateral geniculate nucleus of the cat. Brain Res. 1970, 20: 181-191.View ArticlePubMedGoogle Scholar
- Montero VM, Scott GL: Synaptic terminals in the dorsal lateral geniculate nucleus from neurons of the thalamic reticular nucleus: a light and electron microscope autoradiographic study. Neuroscience. 1981, 6: 2561-2577.View ArticlePubMedGoogle Scholar
- Seabrook TA, Krahe TE, Govindaiah G, Guido W: Interneurons in the mouse visual thalamus maintain a high degree of retinal convergence throughout postnatal development. Neural Dev. 2013, 8: 24-PubMed CentralView ArticlePubMedGoogle Scholar
- Rafols JA, Valverde F: The structure of the dorsal lateral geniculate nucleus in the mouse. A Golgi and electron microscopic study. J Comp Neurol. 1973, 150: 303-332.View ArticlePubMedGoogle Scholar
- Niimi K, Kanaseki T, Takimoto T: The comparative anatomy of the ventral nucleus of the lateral geniculate body in mammals. J Comp Neurol. 1963, 121: 313-323.View ArticlePubMedGoogle Scholar
- Harrington ME: The ventral lateral geniculate nucleus and the intergeniculate leaflet: interrelated structures in the visual and circadian systems. Neurosci Biobehav Rev. 1997, 21: 705-727.View ArticlePubMedGoogle Scholar
- Fox MA, Guido W: Shedding light on class-specific wiring: development of intrinsically photosensitive retinal ganglion cell circuitry. Mol Neurobiol. 2011, 44: 321-329.PubMed CentralView ArticlePubMedGoogle Scholar
- Cosenza RM, Moore RY: Afferent connections of the ventral lateral geniculate nucleus in the rat: an HRP study. Brain Res. 1984, 310: 367-370.View ArticlePubMedGoogle Scholar
- Hattar S, Kumar M, Park A, Tong P, Tung J, Yau KW, Berson DM: Central projections of melanopsin-expressing retinal ganglion cells in the mouse. J Comp Neurol. 2006, 497: 326-349.PubMed CentralView ArticlePubMedGoogle Scholar
- Huberman AD, Manu M, Koch SM, Susman MW, Lutz AB, Ullian EM, Baccus SA, Barres BA: Architecture and activity-mediated refinement of axonal projections from a mosaic of genetically identified retinal ganglion cells. Neuron. 2008, 59: 425-438.View ArticlePubMedGoogle Scholar
- Huberman AD, Wei W, Elstrott J, Stafford BK, Feller MB, Barres BA: Genetic identification of an On-Off direction-selective retinal ganglion cell subtype reveals a layer-specific subcortical map of posterior motion. Neuron. 2009, 62: 327-334.PubMed CentralView ArticlePubMedGoogle Scholar
- Kim IJ, Zhang Y, Yamagata M, Meister M, Sanes JR: Molecular identification of a retinal cell type that responds to upward motion. Nature. 2008, 452: 478-482.View ArticlePubMedGoogle Scholar
- Kim IJ, Zhang Y, Meister M, Sanes JR: Laminar restriction of retinal ganglion cell dendrites and axons: subtype-specific developmental patterns revealed with transgenic markers. J Neurosci. 2010, 30: 1452-1462.PubMed CentralView ArticlePubMedGoogle Scholar
- Kay JN, De la Huerta I, Kim IJ, Zhang Y, Yamagata M, Chu MW, Meister M, Sanes JR: Retinal ganglion cells with distinct directional preferences differ in molecular identity, structure, and central projections. J Neurosci. 2011, 31: 7753-7762.PubMed CentralView ArticlePubMedGoogle Scholar
- Ecker JL, Dumitrescu ON, Wong KY, Alam NM, Chen SK, LeGates T, Renna JM, Prusky GT, Berson DM, Hattar S: Melanopsin-expressing retinal ganglion-cell photoreceptors: cellular diversity and role in pattern vision. Neuron. 2010, 67: 49-60.PubMed CentralView ArticlePubMedGoogle Scholar
- Osterhout JA, Josten N, Yamada J, Pan F, Wu SW, Nguyen PL, Panagiotakos G, Inoue YU, Egusa SF, Volgyi B, Inoue T, Bloomfield SA, Barres BA, Berson DM, Feldheim DA, Huberman AD: Cadherin-6 mediates axon-target matching in a non-image-forming visual circuit. Neuron. 2011, 71: 632-639.PubMed CentralView ArticlePubMedGoogle Scholar
- Triplett JW, Wei W, Gonzalez C, Sweeney NT, Huberman AD, Feller MB, Feldheim DA: Dendritic and axonal targeting patterns of a genetically-specified class of retinal ganglion cells that participate in image-forming circuits. Neural Dev. 2014, 9: 2-PubMed CentralView ArticlePubMedGoogle Scholar
- Sweeney NT, Tierney H, Feldheim DA: Tbr2 is required to generate a neural circuit mediating the pupillary light reflex. J Neurosci. 2014, 34: 5447-5453.PubMed CentralView ArticlePubMedGoogle Scholar
- Jacobs EC, Campagnoni C, Kampf K, Reyes SD, Kalra V, Handley V, Xie YY, Hong-Hu Y, Spreur V, Fisher RS, Campagnoni AT: Visualization of corticofugal projections during early cortical development in a tau-GFP-transgenic mouse. Eur J Neurosci. 2007, 25: 17-30.View ArticlePubMedGoogle Scholar
- Seabrook T, El Danaf R, Krahe TE, Fox MA, Guido W: Retinal input regulates the timing of corticogeniculate innervation. J Neurosci. 2013, 33: 10085-10097.PubMed CentralView ArticlePubMedGoogle Scholar
- Su J, Haner CV, Imbery TE, Brooks JM, Morhardt DR, Gorse K, Guido W, Fox MA: Reelin is required for class-specific retinogeniculate targeting. J Neurosci. 2011, 31: 575-586.PubMed CentralView ArticlePubMedGoogle Scholar
- Fukuda T, Aika Y, Heizmann CW, Kosaka T: GABAergic axon terminals at perisomatic and dendritic inhibitory sites show different immunoreactivities against two GAD isoforms, GAD67 and GAD65, in the mouse hippocampus: a digitized quantitative analysis. J Comp Neurol. 1998, 395: 177-194.View ArticlePubMedGoogle Scholar
- Hardwick C, French SJ, Southam E, Totterdell S: A comparison of possible markers for chandelier cartridges in rat medial prefrontal cortex and hippocampus. Brain Res. 2005, 1031: 238-244.View ArticlePubMedGoogle Scholar
- Fish KN, Sweet RA, Lewis DA: Differential distribution of proteins regulating GABA synthesis and reuptake in axon boutons of subpopulations of cortical interneurons. Cereb Cortex. 2011, 21: 2450-2460.PubMed CentralView ArticlePubMedGoogle Scholar
- de Lima AD, Montero VM, Singer W: The cholinergic innervation of the visual thalamus: an EM immunocytochemical study. Exp Brain Res. 1985, 59: 206-212.View ArticlePubMedGoogle Scholar
- Grant E, Hoerder-Suabedissen A, Molnar Z: Development of the corticothalamic projections. Front Neurosci. 2012, 6: 53-PubMed CentralView ArticlePubMedGoogle Scholar
- Bourassa J, Deschenes M: Corticothalamic projections from the primary visual cortex in rats: a single fiber study using biocytin as an anterograde tracer. Neuroscience. 1995, 66: 253-263.View ArticlePubMedGoogle Scholar
- Wang SW, Kim BS, Ding K, Wang H, Sun D, Johnson RL, Klein WH, Gan L: Requirement for math5 in the development of retinal ganglion cells. Genes Dev. 2001, 15: 24-29.PubMed CentralView ArticlePubMedGoogle Scholar
- Brooks JM, Su J, Levy C, Wang JS, Seabrook TA, Guido W, Fox MA: A molecular mechanism regulating the timing of corticogeniculate innervation. Cell Rep. 2013, 5: 573-581.View ArticlePubMedGoogle Scholar
- Sherman SM, Guillery RW: On the actions that one nerve cell can have on another: distinguishing “drivers” from “modulators”. Proc Natl Acad Sci U S A. 1998, 95: 7121-7126.PubMed CentralView ArticlePubMedGoogle Scholar
- Petrof I, Sherman SM: Functional significance of synaptic terminal size in glutamatergic sensory pathways in thalamus and cortex. J Physiol. 2013, 591: 3125-3131.PubMed CentralView ArticlePubMedGoogle Scholar
- Su J, Klemm MA, Josephson AM, Fox MA: Contributions of VLDLR and LRP8 in the establishment of retinogeniculate projections. Neural Dev. 2013, 8: 11-PubMed CentralView ArticlePubMedGoogle Scholar
- Jaubert-Miazza L, Green E, Lo FS, Bui K, Mills J, Guido W: Structural and functional composition of the developing retinogeniculate pathway in the mouse. Vis Neurosci. 2005, 22: 661-676.View ArticlePubMedGoogle Scholar
- Reese BE: ‘Hidden lamination’ in the dorsal lateral geniculate nucleus: the functional organization of this thalamic region in the rat. Brain Res. 1988, 472: 119-137.View ArticlePubMedGoogle Scholar
- Erzurumlu RS, Jhaveri S, Schneider GE: Distribution of morphologically different retinal axon terminals in the hamster dorsal lateral geniculate nucleus. Brain Res. 1988, 461: 175-181.View ArticlePubMedGoogle Scholar
- Dhande OS, Huberman AD: Retinal ganglion cell maps in the brain: implications for visual processing. Curr Opin Neurobiol. 2014, 24: 133-142.PubMed CentralView ArticlePubMedGoogle Scholar
- Rivlin-Etzion M, Zhou K, Wei W, Elstrott J, Nguyen PL, Barres BA, Huberman AD, Feller MB: Transgenic mice reveal unexpected diversity of on-off direction-selective retinal ganglion cell subtypes and brain structures involved in motion processing. J Neurosci. 2011, 31: 8760-8769.PubMed CentralView ArticlePubMedGoogle Scholar
- Hong YK, Kim IJ, Sanes JR: Stereotyped axonal arbors of retinal ganglion cell subsets in the mouse superior colliculus. J Comp Neurol. 2011, 519: 1691-1711.PubMed CentralView ArticlePubMedGoogle Scholar
- Mize RR, Horner LH: Retinal synapses of the cat medial interlaminar nucleus and ventral lateral geniculate nucleus differ in size and synaptic organization. J Comp Neurol. 1984, 224: 579-590.View ArticlePubMedGoogle Scholar
- Dhande OS, Hua EW, Guh E, Yeh J, Bhatt S, Zhang Y, Ruthazer ES, Feller MB, Crair MC: Development of single retinofugal axon arbors in normal and beta2 knock-out mice. J Neurosci. 2011, 31: 3384-3399.PubMed CentralView ArticlePubMedGoogle Scholar
- Chen C, Regehr WG: Developmental remodeling of the retinogeniculate synapse. Neuron. 2000, 28: 955-966.View ArticlePubMedGoogle Scholar
- Stelzner DJ, Baisden RH, Goodman DC: The ventral lateral geniculate nucleus, pars lateralis of the rat. Synaptic organization and conditions for axonal sprouting. Cell Tissue Res. 1976, 170: 435-454.View ArticlePubMedGoogle Scholar
- Singh R, Su J, Brooks J, Terauchi A, Umemori H, Fox MA: Fibroblast growth factor 22 contributes to the development of retinal nerve terminals in the dorsal lateral geniculate nucleus. Front Mol Neurosci. 2012, 4: 61-PubMed CentralView ArticlePubMedGoogle Scholar
- Guido W: Refinement of the retinogeniculate pathway. J Physiol. 2008, 586: 4357-4362.PubMed CentralView ArticlePubMedGoogle Scholar
- Hong YK, Chen C: Wiring and rewiring of the retinogeniculate synapse. Curr Opin Neurobiol. 2011, 21: 228-237.PubMed CentralView ArticlePubMedGoogle Scholar
- Frost DO: Anomalous visual connections to somatosensory and auditory systems following brain lesions in early life. Brain Res. 1982, 255: 627-635.View ArticlePubMedGoogle Scholar
- Frost DO: Development of anomalous retinal projections to nonvisual thalamic nuclei in Syrian hamsters: a quantitative study. J Comp Neurol. 1986, 252: 95-105.View ArticlePubMedGoogle Scholar
- Campbell G, Frost DO: Target-controlled differentiation of axon terminals and synaptic organization. Proc Natl Acad Sci U S A. 1987, 84: 6929-6933.PubMed CentralView ArticlePubMedGoogle Scholar
- Campbell G, Frost DO: Synaptic organization of anomalous retinal projections to the somatosensory and auditory thalamus: target-controlled morphogenesis of axon terminals and synaptic glomeruli. J Comp Neurol. 1988, 272: 383-408.View ArticlePubMedGoogle Scholar
- Yang Z, Ding K, Pan L, Deng M, Gan L: Math5 determines the competence state of retinal ganglion cell progenitors. Dev Biol. 2003, 264: 240-254.View ArticlePubMedGoogle Scholar
- Varea E, Nacher J, Blasco-Ibanez JM, Gomez-Climent MA, Castillo-Gomez E, Crespo C, Martinez-Guijarro FJ: PSA-NCAM expression in the rat medial prefrontal cortex. Neuroscience. 2005, 136: 435-443.View ArticlePubMedGoogle Scholar
- Xu X, Roby KD, Callaway EM: Mouse cortical inhibitory neuron type that coexpresses somatostatin and calretinin. J Comp Neurol. 2006, 499: 144-160.View ArticlePubMedGoogle Scholar
- Su J, Gorse K, Ramirez F, Fox MA: Collagen XIX is expressed by interneurons and contributes to the formation of hippocampal synapses. J Comp Neurol. 2010, 518: 229-253.PubMed CentralView ArticlePubMedGoogle Scholar
- Voinescu PE, Kay JN, Sanes JR: Birthdays of retinal amacrine cell subtypes are systematically related to their molecular identity and soma position. J Comp Neurol. 2009, 517: 737-750.PubMed CentralView ArticlePubMedGoogle Scholar
- Fortune T, Lurie DI: Chronic low-level lead exposure affects the monoaminergic system in the mouse superior olivary complex. J Comp Neurol. 2009, 513: 542-558.PubMed CentralView ArticlePubMedGoogle Scholar
- Su J, Stenbjorn RS, Gorse K, Su K, Hauser KF, Ricard-Blum S, Pihlajaniemi T, Fox MA: Target-derived matricryptins organize cerebellar synapse formation through alpha3beta1 integrins. Cell Rep. 2012, 2: 223-230.PubMed CentralView ArticlePubMedGoogle Scholar
- Marcucci F, Zou DJ, Firestein S: Sequential onset of presynaptic molecules during olfactory sensory neuron maturation. J Comp Neurol. 2009, 516: 187-198.PubMed CentralView ArticlePubMedGoogle Scholar
- Garbelli R, Inverardi F, Medici V, Amadeo A, Verderio C, Matteoli M, Frassoni C: Heterogeneous expression of SNAP-25 in rat and human brain. J Comp Neurol. 2008, 506: 373-386.View ArticlePubMedGoogle Scholar
- Jakovcevski I, Siering J, Hargus G, Karl N, Hoelters L, Djogo N, Yin S, Zecevic N, Schachner M, Irintchev A: Close homologue of adhesion molecule L1 promotes survival of Purkinje and granule cells and granule cell migration during murine cerebellar development. J Comp Neurol. 2009, 513: 496-510.View ArticlePubMedGoogle Scholar
- Gallart-Palau X, Tarabal O, Casanovas A, Sabado J, Correa FJ, Hereu M, Piedrafita L, Caldero J, Esquerda JE: Neuregulin-1 is concentrated in the postsynaptic subsurface cistern of C-bouton inputs to alpha-motoneurons and altered during motoneuron diseases. Faseb J. 2014, [epub ahead of print]Google Scholar
- Cardona A, Saalfeld S, Schindelin J, Arganda-Carreras I, Preibisch S, Longair M, Tomancak P, Hartenstein V, Douglas RJ: TrakEM2 software for neural circuit reconstruction. PLoS ONE. 2012, 7: e38011-PubMed CentralView ArticlePubMedGoogle Scholar
- Dilger EK, Shin HS, Guido W: Requirements for synaptically evoked plateau potentials in relay cells of the dorsal lateral geniculate nucleus of the mouse. J Physiol. 2011, 589: 919-937.PubMed CentralView ArticlePubMedGoogle Scholar
- Turner JP, Salt TE: Characterization of sensory and corticothalamic excitatory inputs to rat thalamocortical neurons in vitro. J Physiol. 1998, 510 (Pt 3): 829-843.PubMed CentralView ArticlePubMedGoogle Scholar
- Govindaiah , Cox CL: Synaptic activation of metabotropic glutamate receptors regulates dendritic outputs of thalamic interneurons. Neuron. 2004, 41: 611-623.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited.