Cell and Tissue Research

, Volume 316, Issue 1, pp 99–113 | Cite as

Target areas innervated by PACAP-immunoreactive retinal ganglion cells

Regular Article

Abstract

The retinohypothalamic tract (RHT) originates from a subset of retinal ganglion cells (RGCs). The cells of the RHT co-store the neurotransmitters PACAP and glutamate, which in a complex interplay mediate light information to the circadian clock located in the suprachiasmatic nuclei (SCN). These ganglion cells are intrinsically photosensitive probably due to expression of melanopsin, a putative photoreceptor involved in light entrainment. In the present study we examined PACAP-containing retinal projections to the brain using intravitreal injection of the anterograde tracer cholera toxin subunit B (ChB) and double immunostaining for PACAP and ChB. Our results show that the PACAP-containing nerve fibres not only constituted the major projections to the SCN and the intergeniculate leaflet of the thalamus but also had a large terminal field in the olivary pretectal nucleus. The contralateral projection dominated except for the SCN, which showed bilateral innervation. PACAP-containing retinal fibres were also found in the ventrolateral preoptic nucleus, the anterior and lateral hypothalamic area, the subparaventricular zone, the ventral part of the lateral geniculate nucleus and the nucleus of the optic tract. Retinal projections not previously described in the rat also contained PACAP. These new projections were found in the lateral posterior nucleus, the posterior limitans nucleus, the dorsal part of the anterior pretectal nucleus and the posterior and medial pretectal nuclei. Only a few PACAP-containing retinal fibres were found in the superior colliculus. Areas innervated by PACAP-immunoreactive fibres also expressed the PACAP-specific PAC1 receptor as shown by in situ hybridization histochemistry. The findings suggest that PACAP plays a role as neurotransmitter in non-imaging photoperception to target areas in the brain regulating circadian timing, masking, regulation of sleep-wake cycle and pupillary reflex.

Keywords

Cholera toxin subunit B Suprachiasmatic nucleus Entrainment Circadian rhythm Melanopsin Rat (Wistar) 

Abbreviations

3v

Third ventricle

ac

Anterior commissure

AD

Anterodorsal thalamic nucleus

AH

Anterior hypothalamic area

APTD

Anterior pretectal nucleus, dorsal part

ChB

Cholera toxin subunit B

CPu

Caudate putamen

CPT

Commissural pretectal nucleus

DGL

Dorsal geniculate nucleus

IGL

Intergeniculate leaflet

LH

Lateral hypothalamic area

LP

Lateral posterior thalamic nucleus

LS

Lateral septum

MB

Mammillary body

MPO

Medial preoptic nucleus

MPT

Medial pretectal nucleus

oc

Optic chiasma

OPT

Olivary pretectal nucleus

OT

Nucleus of the optic tract

PACAP

Pituitary adenylate cyclase-activating polypeptide

PAC1

PACAP receptor type 1

PAG

Periaqueductal gray

Pe

Periventricular hypothalamic nucleus

PLi

Posterior limitans thalamic nucleus

PPT

Posterior pretectal nucleus

PVT

Paraventricular thalamic nucleus

PVN

Paraventricular hypothalamic nucleus

RGCs

Retinal ganglion cells

RHT

Retinohypothalamic tract

SCN

Suprachiasmatic nucleus

SC

Superior colliculus

SNR

Substantia nigra, reticular part

SON

Supraoptic nucleus

SPVZ

Subparaventricular zone

VGL

Ventral geniculate nucleus

VIP

Vasoactive intestinal peptide

VPAC1

VIP/PACAP receptor type 1

VPAC2

VIP/PACAP receptor type 2

VLPO

Ventrolateral preoptic nucleus

VTA

Ventral tegmental area

Notes

Acknowledgements

The skillful technical assistance of Anita Hansen and Lea Charlotte Larsen is gratefully acknowledged.

References

  1. Angelucci A, Clasca F, Sur M (1996) Anterograde axonal tracing with the subunit B of cholera toxin: a highly sensitive immunohistochemical protocol for revealing fine axonal morphology in adult and neonatal brains. J Neurosci Methods 65:101–112PubMedGoogle Scholar
  2. Belenky MA, Smeraski CA, Provencio I, Sollars PJ, Pickard GE (2003) Melanopsin retinal ganglion cells receive bipolar and amacrine cell synapses. J Comp Neurol 460:380–393CrossRefPubMedGoogle Scholar
  3. Bergström AL, Hannibal J, Hindersson P, Fahrenkrug J (2003) Light-induced phase shift in the Syrian hamster (Mesocricetus auratus) is attenuated by the PACAP receptor antagonist PACAP6–38 or PACAP immunoneutralization. Eur J Neurosci 18:2552–2562CrossRefPubMedGoogle Scholar
  4. Berson DM, Dunn FA, Takao M (2002) Phototransduction by retinal ganglion cells that set the circadian clock. Science 295:1070–1073PubMedGoogle Scholar
  5. Card JP, Moore RY (1989) Organization of lateral geniculate-hypothalamic connections in the rat. J Comp Neurol 284:135–147PubMedGoogle Scholar
  6. Chen D, Buchanan GF, Ding JM, Hannibal J, Gillette MU (1999) PACAP: a pivotal modulator of glutamatergic regulation of the suprachiasmatic circadian clock. Proc Natl Acad Sci U S A 96:13409–13414CrossRefPubMedGoogle Scholar
  7. Clarke RJ, Ikeda H (1985a) Luminance and darkness detectors in the olivary and posterior pretectal nuclei and their relationship to the pupillary light reflex in the rat. I. Studies with steady luminance levels. Exp Brain Res 57:224–232PubMedGoogle Scholar
  8. Clarke RJ, Ikeda H (1985b) Luminance detectors in the olivary pretectal nucleus and their relationship to the pupillary light reflex in the rat. II. Studies using sinusoidal light. Exp Brain Res 59:83–90PubMedGoogle Scholar
  9. Fahrenkrug J, Nielsen HS, Hannibal J (2004) Expression of melanopsin during development of the rat retina. Neuroreport (in press)Google Scholar
  10. Foster RG (2002) Keeping an eye on the time: the Cogan lecture. Invest Ophthalmol Vis Sci 43:1286–1298PubMedGoogle Scholar
  11. Foster RG, Hankins MW (2002) Non-rod, non-cone photoreception in the vertebrates. Prog Retin Eye Res 21:507–527CrossRefPubMedGoogle Scholar
  12. Freedman MS, Lucas RJ, Soni B, von Schantz M, Muñoz M, David-Gray Z, Foster RG (1999) Regulation of mammalian circadian behavior by non-rod, non-cone, ocular photoreceptors. Science 284:502–504PubMedGoogle Scholar
  13. Gooley JJ, Lu J, Chou TC, Scammell TE, Saper CB (2001) Melanopsin in cells of origin of the retinohypothalamic tract. Nat Neurosci 12:1165CrossRefGoogle Scholar
  14. Gooley JJ, Lu J, Fischer D, Saper CB (2003) A broad role for melanopsin in nonvisual photoreception. J Neurosci 23:7093–7106PubMedGoogle Scholar
  15. Hannibal J (2002a) Neurotransmitters of the retino-hypothalamic tract. Cell Tissue Res 309:73–88CrossRefGoogle Scholar
  16. Hannibal J (2002b) Pituitary adenylate cyclase-activating peptide in the rat central nervous system: an immunohistochemical and in situ hybridization study. J Comp Neurol 453:389–417CrossRefPubMedGoogle Scholar
  17. Hannibal J, Fahrenkrug J (2002) Immunoreactive substance P is not part of the retinohypothalamic tract in the rat. Cell Tissue Res 309:293–299CrossRefPubMedGoogle Scholar
  18. Hannibal J, Mikkelsen JD, Clausen H, Holst JJ, Wulff BS, Fahrenkrug J (1995) Gene expression of pituitary adenylate cyclase activating polypeptide (PACAP) in the rat hypothalamus. Regul Pept 55:133–148PubMedGoogle Scholar
  19. Hannibal J, Ding JM, Chen D, Fahrenkrug J, Larsen PJ, Gillette MU, Mikkelsen JD (1997) Pituitary adenylate cyclase activating peptide (PACAP) in the retinohypothalamic tract. A daytime regulator of the biological clock. J Neurosci 17:2637–2644PubMedGoogle Scholar
  20. Hannibal J, Moller M, Ottersen OP, Fahrenkrug J (2000) PACAP and glutamate are co-stored in the retinohypothalamic tract. J Comp Neurol 418:147–155PubMedGoogle Scholar
  21. Hannibal J, Vrang N, Card JP, Fahrenkrug J (2001a) Light dependent induction of c-Fos during subjective day and night in PACAP containing retinal ganglion cells of the retino-hypothalmic tract. J Biol Rhythms 16:457–470PubMedGoogle Scholar
  22. Hannibal J, Brabet P, Jamen F, Nielsen HS, Journot L, Fahrenkrug J (2001b) Dissociation between light induced phase shift of the circadian rhythm and clock gene expression in mice lacking the PACAP type 1 receptor (PAC1). J Neurosci 21:4883–4890Google Scholar
  23. Hannibal J, Hindersson P, Knudsen SM, Georg B, Fahrenkrug J (2002) The photopigment melanopsin is exclusively present in PACAP containing retinal ganglion cells of the retinohypothalamic tract. J Neurosci 22:RC191:1–7Google Scholar
  24. Harmar AJ, Arimura A, Gozes I, Journot L, Laburthe M, Pisegna JR, Rawlings SR, Robberecht P, Said SI, Sreedharan SP, Wank SA, Waschek JA (1998) International Union of Pharmacology. XVIII. Nomenclature of receptors for vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide. Pharmacol Rev 50:265–270PubMedGoogle Scholar
  25. Harrington ME, Hoque S, Hall A, Golombek D, Biello S (1999) Pituitary adenylate cyclase activating peptide phase shifts circadian rhythms in a manner similar to light. J Neurosci 19:6637–6642PubMedGoogle Scholar
  26. Hashimoto H, Nogi H, Mori K, Ohishi H, Shigemoto R, Yamamoto K, Matsuda T, Mizuno N, Nagata S, Baba A (1996) Distribution of the mRNA for a pituitary adenylate cyclase-activating polypeptide receptor in the rat brain: an in situ hybridization study. J Comp Neurol 371:567–577PubMedGoogle Scholar
  27. Hattar S, Liao HW, Takao M, Berson DM, Yau KW (2002) Melanopsin-containing retinal ganglion cells: architecture, projections, and intrinsic photosensitivity. Science 295:1065–1070PubMedGoogle Scholar
  28. Hattar S, Lucas RJ, Mrosovsky N, Thompson S, Douglas RH, Hankins MW, Lem J, Biel M, Hofmann F, Foster RG, Yau KW (2003) Melanopsin and rod-cone photoreceptive systems account for all major accessory visual functions in mice. Nature 424:75–81CrossRefGoogle Scholar
  29. Itaya SK, Van Hoesen GW, Jenq CB (1981) Direct retinal input to the limbic system of the rat. Brain Res 226:33–42CrossRefPubMedGoogle Scholar
  30. Johnson RF, Morin LP, Moore RY (1988) Retinohypothalamic projections in the hamster and rat demonstrated using cholera toxin. Brain Res 462:301–312PubMedGoogle Scholar
  31. Kopp MD, Meissl H, Dehghani F, Korf HW (2001) The pituitary adenylate cyclase-activating polypeptide modulates glutamatergic calcium signalling: investigations on rat suprachiasmatic nucleus neurons. J Neurochem 79:161–171PubMedGoogle Scholar
  32. Legg CR, Cowey A (1977) The role of the ventral lateral geniculate nucleus and posterior thalamus in intensity discrimination in rats. Brain Res 123:261–273CrossRefPubMedGoogle Scholar
  33. Levine JD, Weiss ML, Rosenwasser AM, Miselis RR (1991) Retinohypothalamic tract in the female albino rat: a study using horseradish peroxidase conjugated to cholera toxin. J Comp Neurol 306:344–360PubMedGoogle Scholar
  34. Ling C, Schneider GE, Jhaveri S (1998) Target-specific morphology of retinal axon arbors in the adult hamster. Vis Neurosci 15:559–579CrossRefPubMedGoogle Scholar
  35. Lu J, Shiromani P, Saper CB (1999) Retinal input to the sleep-active ventrolateral preoptic nucleus in the rat. Neuroscience 93:209–214PubMedGoogle Scholar
  36. Lucas RJ, Freedman MS, Munoz M, Garcia-Fernandez JM, Foster RG (1999) Regulation of the mammalian pineal by non-rod, non-cone, ocular photoreceptors. Science 284:505–507PubMedGoogle Scholar
  37. Lucas RJ, Douglas RH, Foster RG (2001) Characterization of an ocular photopigment capable of driving pupillary constriction in mice. Nat Neurosci 4:621–626PubMedGoogle Scholar
  38. Lucas RJ, Hattar S, Takao M, Berson DM, Foster RG, Yau KW (2003) Diminished pupillary light reflex at high irradiances in melanopsin-knockout mice. Science 299:245–247CrossRefPubMedGoogle Scholar
  39. Mikkelsen JD (1992) Visualization of efferent retinal projections by immunohistochemical identification of cholera toxin subunit B. Brain Res Bull 28:619–623PubMedGoogle Scholar
  40. Mombaerts P, Wang F, Dulac C, Chao SK, Nemes A, Mendelsohn M, Edmondson J, Axel R (1996) Visualizing an olfactory sensory map. Cell 87:675–686PubMedGoogle Scholar
  41. Moore RY, Card JP (1994) Intergeniculate leaflet: an anatomically and functionally distinct subdivision of the lateral geniculate complex. J Comp Neurol 344:403–430PubMedGoogle Scholar
  42. Moore RY, Lenn NJ (1972) A retinohypothalamic projection in the rat. J Comp Neurol 146:1–14PubMedGoogle Scholar
  43. Moore RY, Speh JC, Card JP (1995) The retinohypothalamic tract originates from a distinct subset of retinal ganglion cells. J Comp Neurol 352:351–366PubMedGoogle Scholar
  44. Morin LP, Blanchard JH (1997) Neuropeptide Y and enkephalin immunoreactivity in retinorecipient nuclei of the hamster pretectum and thalamus. Vis Neurosci 14:765–777PubMedGoogle Scholar
  45. Morin LP, Blanchard JH, Provencio I (2003) Retinal ganglion cells projections to the hamster suprachiasmatic nucleus, intergeniculate leaflet and visual midbrain: bifurcation and melanopsin immunoreactivity. J Comp Neurol 465:401–416CrossRefPubMedGoogle Scholar
  46. Panda S, Sato TK, Castrucci AM, Rollag MD, DeGrip WJ, Hogenesch JB, Provencio I, Kay SA (2002) Melanopsin (Opn4) requirement for normal light-induced circadian phase shifting. Science 298:2213–2216CrossRefPubMedGoogle Scholar
  47. Panda S, Provencio I, Tu DC, Pires SS, Rollag MD, Castrucci AM, Pletcher MT, Sato TK, Wiltshire T, Andahazy M, Kay SA, Van Gelder RN, Hogenesch JB (2003) Melanopsin is required for non-image-forming photic responses in blind mice. Science 301:525–527CrossRefPubMedGoogle Scholar
  48. Paxinos G, Watson C (1997) The rat brain in stereotaxic coordinates, 3rd edn. Academic Press, San Diego, CAGoogle Scholar
  49. Pickard GE, Smeraski CA, Tomlinson CC, Banfield BW, Kaufman J, Wilcox CL, Enquist LW, Sollars PJ (2002) Intravitreal injection of the attenuated pseudorabies virus PRV Bartha results in infection of the hamster suprachiasmatic nucleus only by retrograde transsynaptic transport via autonomic circuits. J Neurosci 22:2701–2710PubMedGoogle Scholar
  50. Piggins HD, Marchant EG, Goguen D, Rusak B (2001) Phase-shifting effects of pituitary adenylate cyclase activating polypeptide on hamster wheel-running rhythms. Neurosci Lett 305:25–28PubMedGoogle Scholar
  51. Provencio I, Jiang G, De Grip WJ, Hayes WP, Rollag MD (1998) Melanopsin: an opsin in melanophores, brain, and eye. Proc Natl Acad Sci U S A 95:340–345PubMedGoogle Scholar
  52. Provencio I, Rodriguez IR, Jiang G, Hayes WP, Moreira EF, Rollag MD (2000) A novel human opsin in the inner retina. J Neurosci 20:600–605PubMedGoogle Scholar
  53. Provencio I, Rollag MD, Castrucci AM (2002) Photoreceptive net in the mammalian retina. This mesh of cells may explain how some blind mice can still tell day from night. Nature 415:493Google Scholar
  54. Redlin U, Mrosovsky N (1999) Masking by light in hamsters with SCN lesions. J Comp Physiol [A] 184:439–448Google Scholar
  55. Reiner A, Zhang D, Eldred WD (1996) Use of the sensitive anterograde tracer cholera toxin fragment B reveals new details of the central retinal projections in turtles. Brain Behav Evol 48:307–337PubMedGoogle Scholar
  56. Reuss S, Decker K (1997) Anterograde tracing of retinohypothalamic afferents with Fluoro-Gold. Brain Res 745:197–204CrossRefPubMedGoogle Scholar
  57. Riley JN, Card JP, Moore RY (1981) A retinal projection to the lateral hypothalamus in the rat. Cell Tissue Res 214:257–269PubMedGoogle Scholar
  58. Ritter S, Dinh TT (1991) Prior optic nerve transection reduces capsaicin-induced degeneration in rat subcortical visual structures. J Comp Neurol 308:79–90PubMedGoogle Scholar
  59. Ruby NF, Brennan TJ, Xie X, Cao V, Franken P, Heller HC, O’Hara BF (2002) Role of melanopsin in circadian responses to light. Science 298:2211–2213CrossRefPubMedGoogle Scholar
  60. Scalia F (1972) Retinal projections to the olivary pretectal nucleus in the tree shrew and comparison with the rat. Brain Behav Evol 6:237–252PubMedGoogle Scholar
  61. Sefton A, Dreher B (1995) Visual system. In: Paxinos G (ed) The rat nervous system. Academic, New York, pp 833–898Google Scholar
  62. Sheward WJ, Lutz EM, Harmar AJ (1995) The distribution of vasoactive intestinal peptide2 receptor messenger RNA in the rat brain and pituitary gland as assessed by in situ hybridization. Neuroscience 67:409–418CrossRefPubMedGoogle Scholar
  63. Shioda S, Shuto Y, Somogy&vacute, Ari-Vigh A, Legradi G, Onda H, Coy DH, Nakajo S, Arimura A (1997) Localization and gene expression of the receptor for pituitary adenylate cyclase-activating polypeptide in the rat brain. Neurosci Res 28:345–354PubMedGoogle Scholar
  64. Sollars PJ, Smeraski CA, Kaufman JD, Ogilvie MD, Provencio I, Morin LP, Pickard GE (2002) Melanopsin and non-melanopsin expressing retinal ganglion cells innervate the suprachiasmatic nucleus. Soc Neurosci Abstr No.371.21Google Scholar
  65. Trejo LJ, Cicerone CM (1984) Cells in the pretectal olivary nucleus are in the pathway for the direct light reflex of the pupil in the rat. Brain Res 300:49–62CrossRefPubMedGoogle Scholar
  66. Usdin TB, Bonner TI, Mezey E (1994) Two receptors for vasoactive intestinal polypeptide with similar specificity and complementary distribution. Endocrinology 135:2662–2680PubMedGoogle Scholar
  67. Vaudry D, Gonzalez BJ, Basille M, Yon L, Fournier A, Vaudry H (2000) Pituitary adenylate cyclase-activating polypeptide and its receptors: from structure to functions. Pharmacol Rev 52:269–324PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Department of Clinical Biochemistry, Bispebjerg HospitalUniversity of CopenhagenCopenhagenDenmark
  2. 2.Department of Clinical BiochemistryBispebjerg HospitalCopenhagen NVDenmark

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