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In situ hybridization for vasopressin mRNA in the human supraoptic and paraventricular nucleus; quantitative aspects of formalin-fixed paraffin-embedded tissue sections as compared to cryostat sections
Journalof NeuroscienceMethods 64 (1996) 133
Corrigendum
In situ hybridization for vasopressin mRNA in the human s~~~~~c and paraventricular nucleus; quantitative aspects of form&n-fixed paraffin-embedded tissue sections as compared to cryostat sections P.J. Lucassen a**, E. Goudsmit a, C.W. Pool a, G. Mengod b, J.M. Palacios ‘, F.C. R&sheer S.E.F. Guldenaar a, D.F. Swaab a a Gruduatr
School Neurosciences Amsterdam, The Netherlands Institute for Brain Research, Meibergdreef 33, 1105 AZ Amsterdam b Department of Neurochemistry, CID/&SIC, Jordi Girono 1 S-26,08034 Barcelona. Spain ’ Laboratory Almirall, Cardoner 6808024 Barcelona, Spain
a,
20, The Netherlands
Received15 April 1994;revised 8 August 1994;accepted18 August 1994
Abstract In order to study the suitability of formalin-fixed paraffin-embedded brain tissue for vasopressin (AVP)-mRNA detection, we used symmetric halves of 5 human hypothalami. In every case, one half was formalin fixed for IO-35 days and paraffm embedded while the other half was frozen rapidly. Following in situ hybridization (ISH) histochemistry on systematically obtained sections of the supraoptic (SON) and paraventricular nucleus (PVN) of both halves, total amounts of AVP-mRNA in these nuclei were estimated using densitometry of film autoradiographs. Total amounts of radioactivity were found to vary considerably between patients and amounted to 1297 2 302 arbitrary units (AU) (PVN) (mean f SEM) and 2539 f 346 (SON) for the cryostat sections and 868+ 94 (PVN) and 1259 f 126 (SON) for the paraffin tissue.Variations introducedby the method itself yielded a coefficient of variation of only 0,19. Furthermore,a non-significantnegativetrend with postmortemdelay wasfound in cryostattissue,but not in paraffin sections.No effect of fixation time wasobservedin the paraffin tissue.Both ways of tissuetreatmenthave specificadvantages anddisadvantages that may be different for other probesor otherbrain areas.For ISH of a highly abundantmRNA like AVP in a very heterogeneous brainareasuchasthe human hypothalamus, formalin-fixedparaffin-embedded tissue sectionscan be usedfor quantitativeanalysisof entirebrainnucleibecause of the small variation in this tissue,the remarkablygood signal recovery (some75% as comparedto cryostat sections)and its practical advantageswith regardsto anatomicalorientation,storageandsamplingof the tissue. Keywords:
Hypothalamus; In situhybridization; Vasopressin; Formalinfixation;Paraffinembedding; Cryostatsection;Quantitative method
For the above-mentioned article (SSDI 0165 0270(94)00152-9) the corrected addresses are given below. a Graduate School Neurosciences Amsterdam, The Netherlands Institute for Brain Research, Meibergdreef 33, 1105 AZ Amsterdam ZO, The Netherlands.
’ Corresponding author. Tel.: (3 I) 20-566-5500; Fax: (3 1) B-6%- 1006.
0165-0270/%/$12.00 8 1996ElsevierScience B.V. All rightsreserved SSDI 0165-0270(95)00134-4
b Department of Neurochemistry, CID/XC, fordi Girona 18-26, 08034 Barcelona, Spain. ’ Laboratory Almirall, Cardoner 68, 08024 Barcelona, Spain.
Corrigendum
In situ hybridization for vasopressin mRNA in the human s~~~~~c and paraventricular nucleus; quantitative aspects of form&n-fixed paraffin-embedded tissue sections as compared to cryostat sections P.J. Lucassen a**, E. Goudsmit a, C.W. Pool a, G. Mengod b, J.M. Palacios ‘, F.C. R&sheer S.E.F. Guldenaar a, D.F. Swaab a a Gruduatr
School Neurosciences Amsterdam, The Netherlands Institute for Brain Research, Meibergdreef 33, 1105 AZ Amsterdam b Department of Neurochemistry, CID/&SIC, Jordi Girono 1 S-26,08034 Barcelona. Spain ’ Laboratory Almirall, Cardoner 6808024 Barcelona, Spain
a,
20, The Netherlands
Received15 April 1994;revised 8 August 1994;accepted18 August 1994
Abstract In order to study the suitability of formalin-fixed paraffin-embedded brain tissue for vasopressin (AVP)-mRNA detection, we used symmetric halves of 5 human hypothalami. In every case, one half was formalin fixed for IO-35 days and paraffm embedded while the other half was frozen rapidly. Following in situ hybridization (ISH) histochemistry on systematically obtained sections of the supraoptic (SON) and paraventricular nucleus (PVN) of both halves, total amounts of AVP-mRNA in these nuclei were estimated using densitometry of film autoradiographs. Total amounts of radioactivity were found to vary considerably between patients and amounted to 1297 2 302 arbitrary units (AU) (PVN) (mean f SEM) and 2539 f 346 (SON) for the cryostat sections and 868+ 94 (PVN) and 1259 f 126 (SON) for the paraffin tissue.Variations introducedby the method itself yielded a coefficient of variation of only 0,19. Furthermore,a non-significantnegativetrend with postmortemdelay wasfound in cryostattissue,but not in paraffin sections.No effect of fixation time wasobservedin the paraffin tissue.Both ways of tissuetreatmenthave specificadvantages anddisadvantages that may be different for other probesor otherbrain areas.For ISH of a highly abundantmRNA like AVP in a very heterogeneous brainareasuchasthe human hypothalamus, formalin-fixedparaffin-embedded tissue sectionscan be usedfor quantitativeanalysisof entirebrainnucleibecause of the small variation in this tissue,the remarkablygood signal recovery (some75% as comparedto cryostat sections)and its practical advantageswith regardsto anatomicalorientation,storageandsamplingof the tissue. Keywords:
Hypothalamus; In situhybridization; Vasopressin; Formalinfixation;Paraffinembedding; Cryostatsection;Quantitative method
For the above-mentioned article (SSDI 0165 0270(94)00152-9) the corrected addresses are given below. a Graduate School Neurosciences Amsterdam, The Netherlands Institute for Brain Research, Meibergdreef 33, 1105 AZ Amsterdam ZO, The Netherlands.
’ Corresponding author. Tel.: (3 I) 20-566-5500; Fax: (3 1) B-6%- 1006.
0165-0270/%/$12.00 8 1996ElsevierScience B.V. All rightsreserved SSDI 0165-0270(95)00134-4
b Department of Neurochemistry, CID/XC, fordi Girona 18-26, 08034 Barcelona, Spain. ’ Laboratory Almirall, Cardoner 68, 08024 Barcelona, Spain.