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Cloning, Expression, and Localization of MNAR/PELP1 in Rodent Brain: Colocalization in Estrogen Receptor-- But Not in Gonadotropin-Releasing Hormone-Positive Neurons

Institute of Molecular Medicine and Genetics (M.M.K., M.H., C.W., L.M.D.S., K.M.D., V.B.M., D.W.B.), and Institute of Neuroscience (D.W.B.), School of Medicine, Medical College of Georgia, Augusta, Georgia 30912; and Department of Genetics and Stanley S. Scott Cancer Center (R.K.V.), Louisiana State University Health Sciences Center, New Orleans, Louisiana 70112

Address all correspondence and requests for reprints to: Darrell W. Brann, Ph.D., Professor and Associate Director, Institute of Neuroscience, Institute of Molecular Medicine and Genetics, Program in Developmental Neurobiology and Department of Neurology, Medical College of Georgia, 1120 15th Street, Augusta, Georgia 30912. E-mail: dbrann{at}mcg.edu.

MNAR/PELP1 is a recently identified scaffold protein in the human that modulates the nongenomic activity of estrogen receptors by facilitating linkage/cross talk with the Src/Erk activation cascade. We report herein the cloning of rat MNAR/PELP1 and provide new information concerning its distribution in the female rat brain and its degree of colocalization with estrogen receptor- (ER-) and GnRH. PCR-based cloning of MNAR/PELP1 from rat hypothalamus yielded a transcript of approximately 3.4 kb, which shows 86% homology to the published human MNAR/PELP1 sequence and retained all the key binding motifs (PXXP, LXXLL, and glutamic acid clusters) in its primary structure that are known to be critical for its interaction with Src and steroid receptors. RT-PCR revealed that the MNAR/PELP1 transcript is expressed in many regions of the brain, and immunohistochemistry studies showed intense MNAR/PELP1 immunoreactivity (MNAR/PELP1-ir) in areas such as the hypothalamus, cerebral cortex, hippocampus, amygdala, and cerebellum. MNAR/PELP1-ir principally localized in the nucleus, but some cytoplasmic and plasma membrane-associated staining was also observed. MNAR/PELP1-ir was also primarily neuronal, although some localization in glia cells was observed in select brain regions. Colocalization studies revealed that a majority of ER--positive cells in the brain colocalized MNAR/PELP1-ir. In contrast, MNAR/PELP1-ir rarely colocalized in GnRH neurons. In conclusion, the current study provides evidence that MNAR/PELP1 is expressed in key neural tissues of the rat brain that are known targets of steroid action, that its expression is primarily neuronal, and that MNAR/PELP1-ir is strongly colocalized in ER-, but not GnRH neurons in the rodent brain.

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