cDNA microarray analysis after laser microdissection in proliferating islets of partially pancreatectomized mice.
With islet transplantation having grown in popularity since the introduction of the Edmonton protocol, how to secure an unlimited source of islets has become an urgent problem. To resolve this problem, techniques to induce or proliferate islets are urgently required. To achieve this goal, gene expression analysis using a cDNA microarray in islets of partially pancreatectomized mice, in which the remaining islets regenerate and proliferate with insulin secretion and glucose responsiveness, provides us with valuable information. However, those experiments have two critical problems: first, how to selectively collect the regenerating or proliferating islets, and second, the shortage of total RNA extracted from one islet for a microarray analysis. A useful system was thus designed which combined laser microdissection, cDNA amplification by SMART PCR, which can maintain the relative expression profile of transcripts throughout reactions, and a cDNA microarray. Furthermore, this system is expected to contribute to future studies regarding not only islet regeneration but also the function of the islet itself, and this system may also be applicable to many other types of endocrine tissue. In this review, the details of this system are presented and discussed.
Katsuta
Hitoshi
H
Laboratory of Molecular Immunology and Diabetes, Department of Health Sciences, School of Medicine, Kyushu University, Higashi-ku, Fukuoka, Japan. katsuta@shs.kyushu-u.ac.jp
Koyanagi-Katsuta
Rimiko
R
Shiiba
Masafumi
M
Anzai
Keizo
K
Irie
Tarou
T
Aida
Tadateru
T
Akehi
Yuko
Y
Nakano
Masahiko
M
Yasunami
Yohichi
Y
Harada
Mine
M
Nagafuchi
Seiho
S
Ono
Junko
J
Tachikawa
Tetsuhiko
T
D000818:Animals; D049109:Cell Proliferation; D020869:Gene Expression Profiling / Q000379:methods*; D007515:Islets of Langerhans / Q000502:physiology*; D016381:Islets of Langerhans Transplantation; D007834:Lasers; D051379:Mice; D042282:Microdissection; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010180:Pancreatectomy; D012038:Regeneration / Q000235:genetics*; D023822:Tissue Engineering