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Adaptation of seichert's technique to embryonic heart marking
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ADAPTATION OF SEICHERT'S TECHNIQUE TO EM'BRYONIC HEART MARKING. A,Arfinega, J.A. Contreras, L.Alvarez, J.E. IFernandez, F. Revelles. Dept. Ciencias Morifol6gicas, Ftad. Medicina. Univ. Granada. Granada. Spain.
OBlaTION OF i'~:I:L~AL ~ I E S AGAINST[~RYOXEC HEART TIS,NJE. A. Ar~nega, J.E. Fernandez, M.A. [opez, F. Gsrrido and L. Alvarez, Dept. of Morphology, School of Medicine, Univ. of Granada and Dept. of Clinical Analysis, Virgen Nieves Hosp., Granada, Spain.
Marking techniques are undoubtably the most appropriate procedure for studying cell movement during embryonic development. The gelatin - India ink method, used by Sei chert in 1965 and subsequently by other researchers has shown itself to be the most effective in the study of cardiac morphogenesis. The original procedure howerver presentd some drawbacks which almost inervitably led to high mortality rates.The modifications of this nonetheless classic techniQue, developed in our laboratory, include: ~i. Substitution of the traditionally used marking instruments with a newly designed curved carrier. ~. Modification of the technique for taking up the gelatin - India in mixture on the carrier. ~hese changes have consistently led to: ~. Greater ease in the marking procedure. P. Significantly higher survival rates. 8. Improved results.
With hybridema biqtechnology we have produced monoclonal antibodies, overcoming a considerable nunber of obstacle encountered with conventional heteroantisera in the study of membrane proteins. Material and Methods. An i ~ z a t i o n protocol similar to that of Stali ~as used. Over a total'period of 24 days four immunizations were performed in mice, employing minced embryonic hearts (heart loop only) which were centrifuged in order to obtain a pellet which was then combined with 0~2 ml P~S. Immunizations were carried out at 7 day intervals except for the final stimulation which took place only three days after the preceding injection. Cell fusion was performed following a slight modification of Springer,s procedure. The myelc~m cells line used was non Ig-secreting Sp 2/15. Positive hybrids were selected on the basis of positive indirect inmunofluorescence assays in serial cryostatic sections of embryonic heart tissue exposed to developing hybrid supernatant. This technique yielded evidence that moneelonal antibodies had in fact been raised against embryonic heart cells.
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274 EVOLUTIONS OF PREHISTORIC F',UMAN F APULATIONS IN THE IBERIAN PENINSULA. M. C. Boteiia and C. Garcla. Department of N'orphoiogical Sciences. Institute "F.Ol6riz" University o f G r a n a d a , 18012 G r a n a d a Spain.
TK:T EMBRYOLOGICAL COLLECTION OF THE HUBRECHT LABORATORY, G.C. Bangma. Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Uppsalalaan 8, 3584 CT Utrecht. The Hubrecht Laboratory houses the Embryological Collection, an extensive and unique collection of microscopical slides and specimens preserved in alcohol pertaining to the normal development and placentation of more than 600 species of vertebrates. Numerous series of embryonic material of rare and endangered mammalian species are represented, e.g. Marsupials, Insectivores, Primates. The Embryological Collection is expressly available for morphological, descriptive and comparative embryological and placentological studies by research workers in all countries. It may be consulted at the Hubrecht Laboratory on application. Extensive card indexes, catalogues and original notes are available. On certain conditions material of the Collection can be borrowed. A concise catalogue is available on request. In special cases financial support in the form of shortterm maintenance grants is available.
A n t h r o p o i o g i c a l f e a t u r e s o f the vari ous c u l t u r a l periods in the Iberian Penin sula from ~000 B.C. to 1 0 0 0 B.C. ace s t u d i e d . Racial c h a r a c t e r i s t i c s o f popula t i o n s are i n v e s t i g a t e d and m i c r o e v o l u t i o nary f a c t o r s are analyzed. A. large ser i e s o f stadistical parameters correspon-
ding to the Neolithic, Calcoiithic, Lower Bronee Age and Upper Bronce Age have been examined on a region by region as weii as zone by zone basis in order to attempt to generate an overall view of the development of the primitive peopies in the Iberian Peninsula.
96S
272
ADAPTATION OF SEICHERT'S TECHNIQUE TO EM'BRYONIC HEART MARKING. A,Arfinega, J.A. Contreras, L.Alvarez, J.E. IFernandez, F. Revelles. Dept. Ciencias Morifol6gicas, Ftad. Medicina. Univ. Granada. Granada. Spain.
OBlaTION OF i'~:I:L~AL ~ I E S AGAINST[~RYOXEC HEART TIS,NJE. A. Ar~nega, J.E. Fernandez, M.A. [opez, F. Gsrrido and L. Alvarez, Dept. of Morphology, School of Medicine, Univ. of Granada and Dept. of Clinical Analysis, Virgen Nieves Hosp., Granada, Spain.
Marking techniques are undoubtably the most appropriate procedure for studying cell movement during embryonic development. The gelatin - India ink method, used by Sei chert in 1965 and subsequently by other researchers has shown itself to be the most effective in the study of cardiac morphogenesis. The original procedure howerver presentd some drawbacks which almost inervitably led to high mortality rates.The modifications of this nonetheless classic techniQue, developed in our laboratory, include: ~i. Substitution of the traditionally used marking instruments with a newly designed curved carrier. ~. Modification of the technique for taking up the gelatin - India in mixture on the carrier. ~hese changes have consistently led to: ~. Greater ease in the marking procedure. P. Significantly higher survival rates. 8. Improved results.
With hybridema biqtechnology we have produced monoclonal antibodies, overcoming a considerable nunber of obstacle encountered with conventional heteroantisera in the study of membrane proteins. Material and Methods. An i ~ z a t i o n protocol similar to that of Stali ~as used. Over a total'period of 24 days four immunizations were performed in mice, employing minced embryonic hearts (heart loop only) which were centrifuged in order to obtain a pellet which was then combined with 0~2 ml P~S. Immunizations were carried out at 7 day intervals except for the final stimulation which took place only three days after the preceding injection. Cell fusion was performed following a slight modification of Springer,s procedure. The myelc~m cells line used was non Ig-secreting Sp 2/15. Positive hybrids were selected on the basis of positive indirect inmunofluorescence assays in serial cryostatic sections of embryonic heart tissue exposed to developing hybrid supernatant. This technique yielded evidence that moneelonal antibodies had in fact been raised against embryonic heart cells.
273
274 EVOLUTIONS OF PREHISTORIC F',UMAN F APULATIONS IN THE IBERIAN PENINSULA. M. C. Boteiia and C. Garcla. Department of N'orphoiogical Sciences. Institute "F.Ol6riz" University o f G r a n a d a , 18012 G r a n a d a Spain.
TK:T EMBRYOLOGICAL COLLECTION OF THE HUBRECHT LABORATORY, G.C. Bangma. Hubrecht Laboratory, Netherlands Institute for Developmental Biology, Uppsalalaan 8, 3584 CT Utrecht. The Hubrecht Laboratory houses the Embryological Collection, an extensive and unique collection of microscopical slides and specimens preserved in alcohol pertaining to the normal development and placentation of more than 600 species of vertebrates. Numerous series of embryonic material of rare and endangered mammalian species are represented, e.g. Marsupials, Insectivores, Primates. The Embryological Collection is expressly available for morphological, descriptive and comparative embryological and placentological studies by research workers in all countries. It may be consulted at the Hubrecht Laboratory on application. Extensive card indexes, catalogues and original notes are available. On certain conditions material of the Collection can be borrowed. A concise catalogue is available on request. In special cases financial support in the form of shortterm maintenance grants is available.
A n t h r o p o i o g i c a l f e a t u r e s o f the vari ous c u l t u r a l periods in the Iberian Penin sula from ~000 B.C. to 1 0 0 0 B.C. ace s t u d i e d . Racial c h a r a c t e r i s t i c s o f popula t i o n s are i n v e s t i g a t e d and m i c r o e v o l u t i o nary f a c t o r s are analyzed. A. large ser i e s o f stadistical parameters correspon-
ding to the Neolithic, Calcoiithic, Lower Bronee Age and Upper Bronce Age have been examined on a region by region as weii as zone by zone basis in order to attempt to generate an overall view of the development of the primitive peopies in the Iberian Peninsula.
96S