The purification and characterization of plasma membranes and the subcellular distribution of adenylate cyclase in mouse parotid gland

1. 1. Plasma membranes have been purified 17-fold from mouse parotid gland homogenates prepared in hypertonic sucrose media using differential centrifugation. The method is fast and simple. The membranes were characterised by electron microscopy, enzyme composition and chemical composition. Further purification was achieved by isopycnic centrifugation in discontinuous sucrose gradients. 2. 2. The purified membranes contain an adenylate cyclase activity which is stimulated by isoproterenol and fluoride. Only 50% of the total adenylate cyclase activity sedimented in the plasma membrane fraction. The rest of the activity resided in the crude nuclear and mitochondrial pellets. However, this adenylate cyclase activity was not associated with these organelles but with membrane fragments in the pellets. Purified nuclei did not contain adenylate cyclase activity. 3. 3. Adenylate cyclase activity was also localised by electron microscopic cytochemistry. Besides being found at the plasma membra

Adenylate cyclase activity in the parotid gland of the mouse after isoproterenol stimulation.

Previous studies have described a decrease in the activity of adenylate cyclase in the parotid gland of isoproterenol-treated rats. In the present studies, a similar decrease was observed in mice treated with isoproterenol. Studies on the subcellular distribution of adenylate cyclase after isoproterenol stimulation of the parotid gland showed that enzyme activity was increased in the lysosomal fraction and decreased in the cellular membrane fractions. Cytochemical studies on the localization of adenylate cyclase in stimulated gland showed an increase in vesicles which contained enzyme activity and a decrease in activity at the luminal and plasma membranes. It is suggested, based on the present findings and results reported by other investigators, that after isoproterenol stimulation of the parotid gland, adenylate cyclase (along with excess membrane) is degraded by lysosomes. If this suggestion is true, then the observed decrease in adenylate cyclase would have a molecular explanation. </jats:p

New species of <i>Limipolycystis</i> Schilke, 1970 (Rhabdocoela: Kalyptorhynchia: Polycystididae) from the Western Mediterranean

Five new species of Limipolycystis Schilke, 1970 are described and their morphology is compared to that of the only species known, L. curvitubo Schilke, 1970. L. sicilicula n. sp., L. falx n. sp., L. friedae n. sp., L. libra n. sp. and L. wallbergi n. sp. all largely resemble L. curvitubo as to the internal anatomy. They all have a caudally-situated testis and, at least in those species for which good material is available, a seminal receptacle could be seen. This seminal receptacle is tubiform, connecting the bursa with the most proximal part of the oviduct, which is filled with sperm. All these species are mainly distinguishable from each other by details in the shape of the stylet. Diagnoses are provided for all new species and the diagnosis of the genus is emended to reflect the new finds

Coelogynopora Steinbock 1924

Main differences between the species of &lt;i&gt;Coelogynopora&lt;/i&gt; can be derived from the identification key below: &lt;p&gt;1. - Solar organ present.................................................................................... 2&lt;/p&gt; &lt;p&gt;- Solar organ absent.................................................................................... 5&lt;/p&gt; &lt;p&gt; 2.- Male copulatory organ only armed with six to eight small spines 5&ndash;7 &micro;m long.............................. &lt;i&gt;C. visurgis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Male copulatory organ armed with more or larger spines...................................................... 3&lt;/p&gt; &lt;p&gt; 3.- Male copulatory organ armed with six spines 110 &micro;m long............................................... &lt;i&gt;C. solifer&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Male copulatory organ armed with more spines............................................................. 4&lt;/p&gt; &lt;p&gt; 4.- Male copulatory organ armed with nine spines 90&ndash;135 &micro;m long......................................... &lt;i&gt;C. schulzii&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Male copulatory organ armed with 16&ndash;17 spines 70&ndash;80 &micro;m long*..................................... &lt;i&gt;C. multispina&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 5. - Male copulatory organ without hard structures.................................................... &lt;i&gt;C.gynocotyla&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Male copulatory organ with hard structures................................................................. 6&lt;/p&gt; &lt;p&gt;6.- Sclerotic parts of male copulatory organ with central stylet/pipe................................................ 7&lt;/p&gt; &lt;p&gt;- Sclerotic parts of male copulatory organ without central stylet/pipe............................................. 16&lt;/p&gt; &lt;p&gt;7.- Accessory spines present............................................................................... 8&lt;/p&gt; &lt;p&gt;- Accessory spines absent............................................................................... 12&lt;/p&gt; &lt;p&gt; 8.- Accessory spines consisting of one pair of strong spines with hooked tips located caudally of the central spines; central spines fused and with tips oriented towards caudal end.................................................... &lt;i&gt;C. birostrata&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Accessory spines consisting of more than one pair of strong needles............................................. 9&lt;/p&gt; &lt;p&gt;* Armonies (2017) mentions 16 needles in the description, but 17 needles are depicted in figs. 2b and 3a.&lt;/p&gt; &lt;p&gt; 9.- Two groups of three accessory spines present lateral to the central spines, consisting of three different pairs of needles; four or five slightly bent hooks with blunt spur forming a half circle over the central bundle and a small cresent-like piece just in front of central spines............................................................................... &lt;i&gt;C. forcipis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Two groups of more than three accessory spines present...................................................... 10&lt;/p&gt; &lt;p&gt; 10.- Five pairs of identical accessory spines rostral to central spines.................................... &lt;i&gt;C. sopottehlersae&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Five or more pairs of accessory spines organized differently.................................................. 11&lt;/p&gt; &lt;p&gt; 11.- Two groups of six to eight identical accessory spines rostral to central spines, with tips oriented towards caudal end................................................................................................. &lt;i&gt;C. faenofurca&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Two groups of five to six accessory spines of two different types of needles (four to five pairs of fine needles with bent tips and one to two pairs of stronger spines with hooked tip); two short spines on either side alongside the tip of the central stylet................................................................................................ &lt;i&gt;C. cassida&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 12.- Median stylet/pipe with one pair of spines attached basally......................................... &lt;i&gt;C. brachystyla&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- More than one pair of spines present..................................................................... 13&lt;/p&gt; &lt;p&gt; 13.- Median stylet accompanied by two pairs of strong, hook-tipped spines.................................. &lt;i&gt;C. coniuncta&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- More than two pairs of spines present.................................................................... 14&lt;/p&gt; &lt;p&gt; 14.- Median stylet closely associated with two pairs of longer spines (one pair more solid than the other pair) and a second group of 8 shorter spines................................................................................ &lt;i&gt;C. minuta&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Male copulatory organ consisting of 36 needles of three different types fused basally to form a central pipe......... &lt;i&gt;C. alata&lt;/i&gt;&lt;/p&gt; &lt;p&gt;15.- No accessory spines present............................................................................ 16&lt;/p&gt; &lt;p&gt;- Two or more accessory spines present.................................................................... 19&lt;/p&gt; &lt;p&gt; 16.- All spines (26&ndash;28) of central bundle similar, arranged cylindrically...................................... &lt;i&gt;C. gigantea&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Spines of different types present in central bundle........................................................... 17&lt;/p&gt; &lt;p&gt; 17.- Unpaired number of spines in central bundle; 16 (or 18) spines wreath-like arranged around a single central spine..................................................................................................... &lt;i&gt;C. coronata&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Paired number of spines in central bundle................................................................. 18&lt;/p&gt; &lt;p&gt; 18- Central bundle composed of 34 spines arranged in a half circle......................................... &lt;i&gt;C. falcaria&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of 20 spines............................................................ &lt;i&gt;C. frondifera&lt;/i&gt;&lt;/p&gt; &lt;p&gt;19.- Two accessory spines present........................................................................... 20&lt;/p&gt; &lt;p&gt;- More than two accessory spines present.................................................................. 36&lt;/p&gt; &lt;p&gt;20.- Central spines joined basally........................................................................... 21&lt;/p&gt; &lt;p&gt;- Central spines not joined basally........................................................................ 26&lt;/p&gt; &lt;p&gt;21.- Central spines joined basally into one group............................................................... 22&lt;/p&gt; &lt;p&gt; - Central spines joined basally into two groups........................................................ &lt;i&gt;C. nodosa&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 22.- Central bundle of spines pointed caudally............................................................ &lt;i&gt;C. tenuis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Central bundle of spines pointed rostrally................................................................. 23&lt;/p&gt; &lt;p&gt;23.- Accessory spines located laterally from central bundle........................................................24&lt;/p&gt; &lt;p&gt;- Accessory spines located distally from central bundle........................................................ 25&lt;/p&gt; &lt;p&gt; 24.- Central bundle composed of four spines: two fine needles with slightly bent tip centrally and two lateral needles with a hookshaped tip................................................................................... &lt;i&gt;C. sequana&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of six spines: two straight needles and four needles with bent tip................. &lt;i&gt;C. steinboecki&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 25.- Central bundle composed of eight or nine spines: two or three central needles with strongly bent tip and six needles with hooked tip and spur.................................................................................. &lt;i&gt;C. hamulis&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of ten spines: four slender spines, four more robust spines with curved tips and small guard and two short spines associated like forceps............................................................ &lt;i&gt;C. juxtaforcipis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;26.- Accessory spines positioned lateral to central bundle........................................................ 27&lt;/p&gt; &lt;p&gt;- Accessory spines positioned caudal to central bundle........................................................ 28&lt;/p&gt; &lt;p&gt; 27.- Central bundle composed of four different pairs of spines; accessory spines with bent tips; two very small plates just in front of central spines............................................................................ &lt;i&gt;C. faeroernensis&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of five pairs of spines with hooked tips, caudal ends tight together; accessory spines simple, without lateral spur............................................................................... &lt;i&gt;C. sewardensis&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of six pairs of spines of three different types; accessory spines pointed with flared beak-like head........................................................................................ &lt;i&gt;C. poaceaglandis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;28.- Accessory spines pointed caudally....................................................................... 29&lt;/p&gt; &lt;p&gt;- Accessory spines pointed rostally....................................................................... 30&lt;/p&gt; &lt;p&gt; 29.- Accessory spines straight; Central bundle composed of two closely-placed-together groups of spines............ &lt;i&gt;C. scalpri&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Accessory spines with tip bent at right angle towards the middle................................. &lt;i&gt;C. schockaerti&lt;/i&gt; &lt;b&gt;n. sp.&lt;/b&gt;&lt;/p&gt; &lt;p&gt;30.- Tip of accessory spine simple, without appendages.......................................................... 31&lt;/p&gt; &lt;p&gt;- Tip of accessory spines more complex.................................................................... 33&lt;/p&gt; &lt;p&gt; 31.- Central bundle composed of four pairs of simple needles with slightly bent tips.......................... &lt;i&gt;C. tenuiformis&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Central bundle composed of spines of different types........................................................ 32&lt;/p&gt; &lt;p&gt; 32.- Central bundle composed of 11 spines, one central spine accompanied by one pair of fine needles and four pairs of needles with hooked tips; accessory spines with slightly bent tips................................................... &lt;i&gt;C. gallica&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Central bundle composed of 10&ndash;12 spines, six to eight fine, parallel needles, two large cone-shaped spines and two needles positioned diagonally over the cone-shaped spines; accessory spines bent to almost right angles (most of the time)................................................................................................... &lt;i&gt;C. biarmata&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 33.- Central bundle composed of one pair of central spines, with anteriorly another pair of smaller spines........ &lt;i&gt;C. distortofolio&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Central bundle composed of more spines.................................................................. 34&lt;/p&gt; &lt;p&gt; 34.- Accessory spines with strongly bent tip and large appendages on either side, each connected to a muscular bulbus (vesicula granulorum); central bundle composed of five pairs of needles of two different types............................ &lt;i&gt;C. axi&lt;/i&gt;&lt;/p&gt; &lt;p&gt;- Accessory spines more or less straight.................................................................... 35&lt;/p&gt; &lt;p&gt; 35.- Accessory spines with triangular appendix; central bundle composed of four pairs of needles of two different types................................................................................................... &lt;i&gt;C. hymanae&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Accessory spines with strongly bent tip and spur; central bundle composed of six pairs of needles of two different types.............................................................................................. &lt;i&gt;C. hangoensis&lt;/i&gt;&lt;/p&gt; &lt;p&gt; 36.- Four accessory spines present; central bundle composed of one central pair of large spines, surrounded by a circle of about 30 fine needles................................................................................. &lt;i&gt;C. cochleare&lt;/i&gt;&lt;/p&gt; &lt;p&gt; - Two bundles of accessory spines present (12 spines per bundle).......................................... &lt;i&gt;C. erotica&lt;/i&gt;&lt;/p&gt;Published as part of &lt;i&gt;Jouk, Philippe E. H., Revis, Nathalie J. P. &amp; Artois, Tom, 2019, Coelogynopora schockaerti n. sp. (Proseriata: Coelogynoporidae), a remnant of a platyhelminth coldwater fauna in the northwestern Mediterranean?, pp. 409-418 in Zootaxa 4686 (3)&lt;/i&gt; on pages 411-415, DOI: 10.11646/zootaxa.4686.3.6, &lt;a href="http://zenodo.org/record/3998629"&gt;http://zenodo.org/record/3998629&lt;/a&gt