Probe:	probe 1
Probe preparation:	Antisense labelled with S35 DNA
Visualized with:	Autoradiography
Assay notes:	Four oligonucleotide in situ probes were described - Probe 1 (MGI:3817930), Probe 2 (MGI:3817931), Probe 3 (MGI:3817932), and Probe 4 (MGI:3817933). However, the probe used in the figures was not detailed. The four probesgave the same hybridization pattern on several adjacent adult brain, embryonic and fetal serial tissue sections. Sections were fixed in 2% paraformaldehyde for 30 min. The probe used in this assay was 3' labelled.

Structure	Level	Pattern	Image	Note
TS15: neural tube lateral wall	Present	Regionally restricted		Expression was detected in the ventral basal plate of the neural tube.

Structure	Level	Pattern	Image	Note
TS18: future spinal cord	Strong	Regionally restricted		Expression was strong in the future ventral horns at the rostral end of the primitive spinal cord.
TS18: hindbrain mantle layer	Present	Regionally restricted		Expression was detected in the external zone of the mantle layer in the caudal part of the hindbrain.
TS18: hindbrain ventricular layer	Absent
TS18: diencephalon	Weak
TS18: midbrain floor plate	Weak
TS18: sympathetic ganglion	Present

Structure	Level	Pattern	Image	Note
TS19: brain	Present	Regionally restricted		Expression was detected in differentiating neurons.
TS19: future spinal cord	Present	Regionally restricted		Expression was detected in differentiating neurons.
TS19: acoustic ganglion	Present
TS19: facial ganglion	Present
TS19: glossopharyngeal ganglion	Present
TS19: trigeminal ganglion	Present
TS19: vagus ganglion	Present

Structure	Level	Pattern	Image	Note
TS20: hypothalamus	Present		2A
TS20: medulla oblongata	Present		2A
TS20: thalamus	Present		2A
TS20: spinal cord	Present	Regionally restricted	2A	Expression was detected in the anterior, firstly differentiated part of the spinal cord. The posterior part was not labeled.
TS20: olfactory epithelium	Present	Regionally restricted	2A	Expression was detected in the cells located just beneath the superficial layer of the olfactory epithelium.
TS20: dorsal root ganglion	Present	Regionally restricted	2A	Expression was detected in the dorsal root ganglia sensory neurons.
TS20: pons	Present	Regionally restricted	2A	Expression was detected in the basal plate of the pons.
TS20: sympathetic ganglion	Strong	Regionally restricted	2A	Expression was detected in the anlage of the cervical sympathetic ganglia.
TS20: midbrain	Present	Regionally restricted	2A	Expression was detected in the ventral tegmentum nucleus, the ventral tegmental area and the substantia nigra. The tectum was not labeled.
TS20: caudal ganglionic eminence	Weak	Regionally restricted	2A	The ganglionic eminence was faintly labeled in its most external and caudal parts.
TS20: ganglionic eminence	Weak	Regionally restricted	2A	The ganglionic eminence was faintly labeled in its most external and caudal parts.
TS20: medial ganglionic eminence	Absent		2A
TS20: midbrain roof plate	Present	Regionally restricted	2A	Expression was detected in the roof of the midbrain in the outer superficial part of the mantle layer.
TS20: cerebral cortex	Absent		2A	The roof of the neopallial cortex was not labeled.
TS20: cerebellum primordium	Absent		2A	The rhombic lip was not labeled.

Structure	Level	Pattern	Image	Note
TS20: enteric nervous system	Present	Regionally restricted		Expression was detected in the post-migratory cells of the enteric nervous system in the upper part of the gut.

Structure	Level	Pattern	Image	Note
TS21: corpus striatum	Present		2C
TS21: midbrain roof plate	Present		2C
TS21: telencephalon	Present	Regionally restricted	2C	Expression was detected in the roof of the telencephalic vesicles.
TS21: cerebral cortex	Present	Regionally restricted	2C	Expression was detected in the superficial nuclear layer of the telencephalic neopallial cortex.
TS21: olfactory cortex	Present		2C
TS21: olfactory lobe	Present	Regionally restricted	2C	Expression was detected in the olfactory bulb and the tubercle.

Structure	Level	Pattern	Image	Note
TS21: corpus striatum	Present		2D
TS21: midbrain roof plate	Present		2D
TS21: telencephalon	Present	Regionally restricted	2D	Expression was detected in the roof of the telencephalic vesicles.
TS21: cerebral cortex	Present	Regionally restricted	2D	Expression was detected in the superficial nuclear layer of the telencephalic neopallial cortex.
TS21: olfactory cortex	Present		2D
TS21: olfactory lobe	Present	Regionally restricted	2D	Expression was detected in the olfactory bulb and the tubercle.

Structure	Level	Pattern	Image	Note
TS22: cerebellum	Present		2F
TS22: epithalamus	Present		2F
TS22: choroid plexus	Absent		2F
TS22: corpus striatum	Present	Regionally restricted	2F	The corpus striatum laterale is labeled, while the corpus striatum mediale is not labeled.

Structure	Level	Pattern	Image	Note
TS22: dorsal root ganglion	Present		3A

Structure	Level	Pattern	Image	Note
TS21: gland	Present	Regionally restricted	3C	Expression was detected in the adrenal chromaffin cells.
TS21: metanephros	Absent		3C

Structure	Level	Pattern	Image	Note
TS23: enteric nervous system	Present	Regionally restricted	3E	Expression was detected in the parasympathetic myenteric ganglia surrounding the small intestine.

Structure	Level	Pattern	Image	Note
TS21: thoracic ganglion	Present	Regionally restricted	3G	Expression was detected in the inferior sympathetic ganglia.

Structure	Level	Pattern	Image	Note
TS21: cerebellum	Present	Regionally restricted		Expression was detected in the differentiating cells of the cerebellum.
TS21: neural retinal epithelium	Strong	Regionally restricted		The neural retina displayed a strong signal in the innermost layer.
TS21: cochlear ganglion	Strong			Expression was strong in the spiral ganglion.

Structure	Level	Pattern	Image	Note
TS22: retina	Present	Regionally restricted	4A	Only the ganglionic cells of the retina were labeled.

Structure	Level	Pattern	Image	Note
TS26: retina	Present	Regionally restricted	4C	Expression was detected in the differentiated retinal ganglionic cells.

Structure	Level	Pattern	Image	Note
TS28: photoreceptor layer	Present		4E
TS28: photoreceptor layer outer segment	Present		4F

Structure	Level	Pattern	Image	Note
TS28: retina	Present	Regionally restricted		Expression was detected in photoreceptor cells. Bipolar, ganglionic and horizontal cells express faint hybridization signals, if any.

Structure	Level	Pattern	Image	Note
TS23: cochlear ganglion	Present		5A	Ganglionic cells of the spiral ganglion are significantly labeled.

Structure	Level	Pattern	Image	Note
TS23: paraganglion of Zuckerkandl	Strong
TS23: aorta	Present	Regionally restricted		Expression was detected in the media of the aortic wall.
TS23: enteric nervous system	Present	Regionally restricted		Expression was detected in the myenteric plexus along the entire axis of the gut.

Structure	Level	Pattern	Image	Note
TS24: spinal cord	Strong			Expression was strong in the whole spinal cord.
TS24: cranial ganglion	Strong
TS24: medulla oblongata	Strong
TS24: pons	Strong
TS24: hypothalamus	Present	Regionally restricted		Expression was detected in several hypothalamic and thalamic nuclei.
TS24: thalamus	Present	Regionally restricted		Expression was detected in several hypothalamic and thalamic nuclei.
TS24: cerebellum	Present	Regionally restricted		Expression was detected in the deep cerebellar nuclei.
TS24: midbrain roof plate	Present
TS24: telencephalon	Present	Regionally restricted		Expression was detected in the roof of the telencephalic vesicles.
TS24: cerebral cortex	Present			Expression was detected in the neopallial cortex.
TS24: olfactory cortex	Strong	Regionally restricted		Expression was detected in the olfactory cortex and the olfactory tubercle.
TS24: main olfactory bulb	Strong
TS24: spinal ganglion	Strong
TS24: dorsal root ganglion	Present
TS24: sympathetic ganglion	Present
TS24: adrenal gland	Present	Regionally restricted		Expression was detected in the chromaffin cells of the adrenal.
TS24: autonomic ganglion	Present	Regionally restricted		Expression was detected in the parasympathetic ganglia.

Structure	Level	Pattern	Image	Note
TS25: brain	Very strong
TS25: cerebral cortex	Very strong
TS25: hippocampus	Very strong
TS25: hypothalamus	Very strong	Regionally restricted		Expression was very strong in several hypothalamic nuclei and some thalamic nuclei, such as the lateral geniculate nuclei. Significant expression was noted in the midline thalamic nuclei.
TS25: thalamus	Very strong	Regionally restricted		Expression was very strong in several hypothalamic nuclei and some thalamic nuclei, such as the lateral geniculate nuclei. Significant expression was noted in the midline thalamic nuclei.
TS25: brainstem	Present	Regionally restricted		Expression was detected in the dorsal tegmental nuclei, the cranial nerve nuclei, the dorsal and ventral cochlear nuclei and the nuclei of the lateral lemniscus area.
TS25: dorsal root ganglion	Present
TS25: sympathetic ganglion	Present
TS25: vomeronasal organ	Present
TS25: myenteric nerve plexus	Present			Expression was detected in the myenteric ganglia.
TS25: embryo mesenchyme	Present	Regionally restricted		Expression was detected in the ectomesenchymal cells forming connective tissue.

Structure	Level	Pattern	Image	Note
TS26: nasal cavity	Present	Regionally restricted	5C	Expression was detected just beneath the olfactory epithelium surrounding the nasal cavity.

Structure	Level	Pattern	Image	Note
TS26: olfactory cortex	Present	Regionally restricted	5E	Expression was detected in the differentiating olfactory cortex.
TS26: olfactory cortex ventricular layer	Absent		5E

Structure	Level	Pattern	Image	Note
TS26: central nervous system	Strong
TS26: sympathetic ganglion	Present
TS26: autonomic ganglion	Present	Regionally restricted		Expression was detected in parasympathetic cardiac ganglia but was fading in the parasympathetic ganglia of the gut.
TS26: dorsal root ganglion	Weak			Expression was fading in the dorsal root ganglion.

Structure	Level	Pattern	Image	Note
TS21: ovary	Absent

Structure	Level	Pattern	Image	Note
TS21: testis	Absent

Structure	Level	Pattern	Image	Note
TS26: ovary	Absent

Structure	Level	Pattern	Image	Note
TS26: testis	Absent

Structure	Level	Pattern	Image	Note
TS25: autonomic ganglion	Strong	Regionally restricted	6A	Expression was detected in the parasympathetic post-ganglionic neurons of the heart (cardiac ganglia).

Structure	Level	Pattern	Image	Note
TS25: smooth muscle tissue	Strong	Regionally restricted	6C	Expression was detected in the smooth muscle of the truncoconal septa.

Structure	Level	Pattern	Image	Note
TS28: pituitary gland	Present	Regionally restricted	6E	Expression was detected in the anterior pituitary gland.

Structure	Level	Pattern	Image	Note
TS28: islets of Langerhans	Present
TS28: cerebral cortex	Present
TS28: dentate gyrus	Present
TS28: hippocampus	Present
TS28: hypothalamic nucleus	Present	Regionally restricted		Expression was detected in several hypothalamic nuclei.
TS28: cerebellum Purkinje cell layer	Present
TS28: lateral septal nucleus	Present
TS28: substantia nigra pars compacta	Present
TS28: ventral tegmental area	Present
TS28: telencephalon septum	Present	Regionally restricted		Expression was detected in the lateral and dorsal septal nuclei.
TS28: corpus callosum	Absent
TS28: corpus striatum	Absent
TS28: thalamus	Absent
TS28: white matter	Absent

Structure	Level	Pattern	Image	Note
TS28: olfactory cortex	Present		7A	Expression was detected in all neurons of the primary olfactory cortex.

Structure	Level	Pattern	Image	Note
TS28: cingulate cortex	Present		7B

Structure	Level	Pattern	Image	Note
TS28: subiculum	Present	Regionally restricted	7C	Expression was detected in both interneurons and pyramidal cells of the subiculum.

Structure	Level	Pattern	Image	Note
TS28: medial habenular nucleus	Present		7D

Structure	Level	Pattern	Image	Note
TS28: area postrema	Present		7E
TS28: subfornical organ	Present		7E

Structure	Level	Pattern	Image	Note
TS28: cerebellum Purkinje cell layer	Present		7F

Structure	Level	Pattern	Image	Note
TS28: arcuate nucleus	Present		7G

Structure	Level	Pattern	Image	Note
TS28: medial preoptic nucleus	Present		7H

Structure	Level	Pattern	Image	Note
TS28: dorsomedial hypothalamic nucleus	Present		7I
TS28: paraventricular hypothalamic nucleus	Present		7I

Structure	Level	Pattern	Image	Note
TS28: cerebellar cortex	Present
TS28: inferior olivary nucleus	Present
TS28: hippocampus pyramidal cell layer	Present
TS28: hippocampus	Present	Regionally restricted		Expression was detected in both the pyramidal and basket cells of the hippocampus.
TS28: dentate gyrus granule cell layer	Strong
TS28: corpus striatum	Absent
TS28: thalamus	Present	Regionally restricted		Expression was detected in the midline thalamic nuclei.
TS28: interpeduncular nucleus	Present
TS28: substantia nigra pars compacta	Present
TS28: ventral tegmental area	Present
TS28: ventromedial hypothalamic nucleus	Present
TS28: hypothalamic nucleus	Present	Regionally restricted		Expression was detected in the preoptic periventricular hypothalamic nucleus, the ventral premammillary nucleus and the posterior periventricular nucleus.
TS28: amygdala	Present	Regionally restricted		Several nuclei belonging to the extended amygdala showed expression.
TS28: dorsal raphe nucleus	Present	Regionally restricted		Expression was detected in the serotoninergic neurons of the dorsal raphe nucleus.
TS28: midbrain	Present	Regionally restricted		Expression was detected in the midbrain median raphe nucleus.
TS28: hindbrain	Present	Regionally restricted		Expression was detected in the caudal linear raphe nucleus.