Immunohistochemical and in situ mRNA hybridisation techniques to determine the distribution of ion channels in human brain: a study of neuronal voltage-dependent calcium channels

• Published in: Brain research. Brain research protocols Vol. 1; no. 3; pp. 299 - 306
• Main Authors: McCormack, Alison L, Day, Nicola C, Craig, Peter J, Smith, William, Beattie, Ruth E, Volsen, Stephen G
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.08.1997
• Subjects: Autoradiography; Brain - cytology; Brain - metabolism; Calcium channel; Calcium Channels - genetics; Calcium Channels - metabolism; Calcium Channels - physiology; Electrophysiology; Human brain; Humans; Immunohistochemistry; Immunohistochemistry - methods; In Situ Hybridization - methods; mRNA hybridisation, in situ; Neurons - metabolism; RNA, Messenger - metabolism; Tissue Distribution; Immunohistochemistry; Human brain; mRNA hybridisation, in situ; Calcium channel
• ISSN: 1385-299X
• DOI: 10.1016/S1385-299X(97)00003-2

The molecular, structural and functional characterisation of ion channels in the CNS forms an area of intense investigation in current brain research. For strategic and logistical reasons, rodents have historically been the species of choice for these studies. The examination of human CNS tissues generally presents the investigator with specific challenges that are often less problematic in animal studies, e.g. post-mortem delay/agonal status, and thus both the experimental design and techniques must be manipulated accordingly. Since much pharmaceutical interest is currently focused on neuronal ion channels, the examination of their expression in human brain material is of particular importance. We describe here the details of methods that we have developed and used successfully in the study of the expression of voltage-dependent calcium channels (VDCCs) in human CNS tissues [15]. Presynaptic neuronal VDCCs control neurotransmitter release [11, 8]and are important new drug targets [3]. They are composed of three subunits, α 1, β and α 2/ δ [4]and multiple gene classes of each protein have been identified [7, 12, 14]. Little is known, however, about the distribution of neuronal VDCCs in the human central nervous system, although initial studies have been performed in rat and rabbit [6, 13, 16].