Single-cell RT-PCR, in situ hybridization histochemical, and immunohistochemical studies of substance P and enkephalin co-occurrence in striatal projection neurons in rats

H. B. Wang, A. V. Laverghetta, Robert Foehring, Yunping Deng, Z. Sun, K. Yamamoto, W. L. Lei, Y. Jiao, Anton Reiner

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Abstract

Single-cell RT-PCR studies in 3-4-week-old rats have raised the possibility that as many as 20% of striatal projection neurons may be a unique type that contains both substance P (SP) and enkephalin (ENK). We used single-cell RT-PCR, retrograde labeling, in situ hybridization histochemistry, and immunolabeling to characterize the abundance of this cell type, its projection target(s), and any developmental changes in its frequency. We found by RT-PCR that 11% of neurons containing either SP or ENK contained both in 4-week-old rats, while in 4-month-old rats SP/ENK colocalization was only 3%. SP-only neurons tended to co-contain dynorphin and ENK-only neurons neurotensin, while SP/ENK neurons tended to contain dynorphin. Single-cell RT-PCR showed SP/ENK co-occurrence in 4-week-old rats to be no more common among striatal neurons retrogradely labeled from the substantia nigra than among those retrogradely labeled from globus pallidus. Double-label in situ hybridization showed SP/ENK perikarya to be scattered throughout striatum, making up 8% of neurons containing either SP or ENK at 4 weeks, but only 4% at 4 months. Immunolabeling showed that presumptive striatal terminals in globus pallidus externus, globus pallidus internus and substantia nigra pars reticulata that colocalized SP and ENK were scarce. Terminals colocalizing SP and ENK were, however, abundant in the substantia nigra pars compacta. Thus, SP-only and ENK-only neurons make up the vast majority of striatal projection neurons in rats, the frequency of SP/ENK colocalizing striatal neurons is low in adult rats (3-4%), and SP/ENK colocalizing neurons primarily project to SNc but do not appear to be confined to striosomes.

Original languageEnglish (US)
Pages (from-to)178-199
Number of pages22
JournalJournal of Chemical Neuroanatomy
Volume31
Issue number3
DOIs
StatePublished - Apr 1 2006

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Corpus Striatum
Enkephalins
Substance P
In Situ Hybridization
Neurons
Polymerase Chain Reaction
Globus Pallidus
Dynorphins
Neurotensin
Substantia Nigra

All Science Journal Classification (ASJC) codes

  • Cellular and Molecular Neuroscience

Cite this

Single-cell RT-PCR, in situ hybridization histochemical, and immunohistochemical studies of substance P and enkephalin co-occurrence in striatal projection neurons in rats. / Wang, H. B.; Laverghetta, A. V.; Foehring, Robert; Deng, Yunping; Sun, Z.; Yamamoto, K.; Lei, W. L.; Jiao, Y.; Reiner, Anton.

In: Journal of Chemical Neuroanatomy, Vol. 31, No. 3, 01.04.2006, p. 178-199.

Research output: Contribution to journalArticle

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abstract = "Single-cell RT-PCR studies in 3-4-week-old rats have raised the possibility that as many as 20{\%} of striatal projection neurons may be a unique type that contains both substance P (SP) and enkephalin (ENK). We used single-cell RT-PCR, retrograde labeling, in situ hybridization histochemistry, and immunolabeling to characterize the abundance of this cell type, its projection target(s), and any developmental changes in its frequency. We found by RT-PCR that 11{\%} of neurons containing either SP or ENK contained both in 4-week-old rats, while in 4-month-old rats SP/ENK colocalization was only 3{\%}. SP-only neurons tended to co-contain dynorphin and ENK-only neurons neurotensin, while SP/ENK neurons tended to contain dynorphin. Single-cell RT-PCR showed SP/ENK co-occurrence in 4-week-old rats to be no more common among striatal neurons retrogradely labeled from the substantia nigra than among those retrogradely labeled from globus pallidus. Double-label in situ hybridization showed SP/ENK perikarya to be scattered throughout striatum, making up 8{\%} of neurons containing either SP or ENK at 4 weeks, but only 4{\%} at 4 months. Immunolabeling showed that presumptive striatal terminals in globus pallidus externus, globus pallidus internus and substantia nigra pars reticulata that colocalized SP and ENK were scarce. Terminals colocalizing SP and ENK were, however, abundant in the substantia nigra pars compacta. Thus, SP-only and ENK-only neurons make up the vast majority of striatal projection neurons in rats, the frequency of SP/ENK colocalizing striatal neurons is low in adult rats (3-4{\%}), and SP/ENK colocalizing neurons primarily project to SNc but do not appear to be confined to striosomes.",
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AU - Laverghetta, A. V.

AU - Foehring, Robert

AU - Deng, Yunping

AU - Sun, Z.

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AU - Lei, W. L.

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AU - Reiner, Anton

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