Is life conceivable Is life conceivable without without chemotaxis ?chemotaxis ?

Dr. habil. Kőhidai László2011.

Trichinella spiralis (1)Trichinella spiralis (1)

+ +20 - 80

200 - 400

+ +

<

Trichinella (2)Trichinella (2)

+ +

+

Trichinella (3)Trichinella (3)

Insects (1)Insects (1)

Responses to pheromones: in sec.-sResponses to pheromones: in sec.-s- direction of flying- direction of flying- speed- speed- pattern (zick-zack)- pattern (zick-zack)

Modulation of odorant receptors is durable Modulation of odorant receptors is durable 1-2 min. - 20-95 min.1-2 min. - 20-95 min.

ipsenol, ipsdienol, cis-verbenolipsenol, ipsdienol, cis-verbenol

Dose ranges of responses are wideDose ranges of responses are wide

Insects (2)Insects (2)

Heat-dependent responsesHeat-dependent responses

20 20 ooCC

26 26 ooCC

+steepness of flyingsteepness of flying

Different types of migrations/flying:Different types of migrations/flying:

schemakinezisschemakinezis zikk-zakkzikk-zakktropotaxistropotaxis look-leaplook-leapklinotaxisklinotaxis

Insects (3)Insects (3)

CockroachCockroach

pheromonespheromones inducing inducing aggregationaggregation

ammónia, methylamine, di-, trimethylamineammónia, methylamine, di-, trimethylamine

1-dimethylamino-2-methyl-2-propanol 1-dimethylamino-2-methyl-2-propanol effective ineffective in 10 pM10 pM(50-1000x more active then other substances)(50-1000x more active then other substances)

Differences in responses to Differences in responses to diffusediffuse and and surface associated surface associated pheromonespheromones

Effect of gamonesEffect of gamones

Androgamone II.Androgamone II.

Androgamone I.Androgamone I.

Gynogamone I.Gynogamone I.

Gynogamone IIGynogamone II..

CoagulationCoagulation

HyaluronidaseHyaluronidase

Parts of female sexual organs releasing Parts of female sexual organs releasing chemoattractantschemoattractants

Cervical mucusCervical mucus

Cavity of uterusCavity of uterus

Follicular fluideFollicular fluide

Cumulus cellsCumulus cells

Intact, still not fertilized oocyteIntact, still not fertilized oocyte

Components of follicular fluideComponents of follicular fluide

Composition is sexual cycle dependantComposition is sexual cycle dependant LHLH Progesterone (1-100 Progesterone (1-100 g/ml)g/ml) Adrenalin (0.001 mg/mllAdrenalin (0.001 mg/mll Oxytocin (0.01 U/ml)Oxytocin (0.01 U/ml) Insulin (repellens)Insulin (repellens) KallikreinKallikrein Anti-thrombin III. Anti-thrombin III.

= spermium receptor= spermium receptor Atrial natriuretic peptide (ANP=ANF)Atrial natriuretic peptide (ANP=ANF) 1000 Da, heat-stable protein1000 Da, heat-stable protein

Other factors synthesized by the female organs Other factors synthesized by the female organs and influencing the chemotaxis and influencing the chemotaxis

pHpH

some proteasessome proteases

resact – Caresact – Ca2+2+ [mM] [mM]

speract – guanylate cyclasesperact – guanylate cyclase

chemoattractant substance perelased from chemoattractant substance perelased from

the vegetative pole of oocytethe vegetative pole of oocyte

Brown algaeBrown algae Ectocarpene 0.89 – 8.9 nmol/lEctocarpene 0.89 – 8.9 nmol/l

the the --electone-distribution is essentialelectone-distribution is essential

Spermiums and chemotaxis (1) Spermiums and chemotaxis (1)

Population of spermiums is rather Population of spermiums is rather heterogeneousheterogeneous

The respiratory response induced by follicular fluide: The respiratory response induced by follicular fluide:

70 % positive70 % positive !!!!!!20 % negative20 % negative

Responsiveness is changing by the age of cells:Responsiveness is changing by the age of cells:

earlyearly maturedmatured agedaged

only these cells express normal responsivenessonly these cells express normal responsiveness

Spermiums and chemotaxis (2) Spermiums and chemotaxis (2)

CaCa2+2+ has a role in: has a role in: chemotaxischemotaxiscell respiratory proc. cell respiratory proc. acrosome reactionacrosome reactioncortocal reactioncortocal reactionfertiléizationfertiléizationcell adhesioncell adhesion

Calmodulin – NAD-kinase – exocytosisCalmodulin – NAD-kinase – exocytosisphosphatases, phosphorylationphosphatases, phosphorylation

guanylate cyclase cGMPguanylate cyclase cGMP

cGMP-depend. CacGMP-depend. Ca2+2+ channel channelCaCa2+2+ influx when spermium reaches the oocyte influx when spermium reaches the oocyte

Spermiums and chemotaxis (3) Spermiums and chemotaxis (3)

Methylation systemMethylation system

tailtail – protein carboxyl-methylase – protein carboxyl-methylasehead – tailhead – tail – MAP – MAP

decreased phospholipid methylation (min. 40%)decreased phospholipid methylation (min. 40%)

cAMP-dependent phosphorylationcAMP-dependent phosphorylation

Protein kinase inhibitors are blockers of the system Protein kinase inhibitors are blockers of the system (42 kD kináz)(42 kD kináz)

Spermiums and chemotaxis (4) Spermiums and chemotaxis (4)

Common receptor-gene family in Common receptor-gene family in odorantodorant receptors receptors and in and in spermiumspermium

fMLF and BOC-fMLPfMLF and BOC-fMLP (10 (10-9-9 – 10 – 10-8-8 M) M) works as chworks as chemoattractantemoattractant not only in leukocytes not only in leukocytes BUT in spermiums, too. BUT in spermiums, too. (leukocyte accumulation is induced in the female (leukocyte accumulation is induced in the female

genital tract)genital tract)

p-nitrophenyl-glycerolp-nitrophenyl-glycerol (PNPG) 10 (PNPG) 10-5-5 M is M is repellentrepellent in spermiums – contraceptive applicationsin spermiums – contraceptive applications

Characterization of pheromonesCharacterization of pheromones

Work on the individuals of the Work on the individuals of the same speciessame species Influence the Influence the sexual behavioursexual behaviour Effects are expressed via pheromone-Effects are expressed via pheromone-receptorsreceptors signaling is signaling is G-protein-linkedG-protein-linked Influence development of hierarchy in the populationInfluence development of hierarchy in the population

qualitativequalitativedifferencesdifferences

quantitativequantitative

Excreted in: Excreted in: feaces, urine, sweet and other body-fluidsfeaces, urine, sweet and other body-fluids Determined by Determined by MHC-genesMHC-genes

the same the same H2H2 (male-female) prefer each other (male-female) prefer each other

Pheromones (2) Pheromones (2)

MiceMice

Estrus-inhibitionEstrus-inhibition

Anestrus Estrus (sensible period lasts 5 days,Anestrus Estrus (sensible period lasts 5 days,but mating desensibilizes it)but mating desensibilizes it)

Non-self male excretes pheromonesNon-self male excretes pheromonesin the in the urineurine

Pheromones (3) Pheromones (3)

Level of heterozygocy Level of heterozygocy is increasedis increased

Pheromones in lower levels of phylogeny (1)Pheromones in lower levels of phylogeny (1)

7 transmembrane receptor7 transmembrane receptor G-protein-linkedG-protein-linked Clathrin haevy-chain dependent internalizationClathrin haevy-chain dependent internalization

Tetrahymena pyriformisTetrahymena pyriformis

Attractant: Attractant: tricosenetricosene ; imprinting ; imprinting ++

Repellent: Repellent: bornyl acetatebornyl acetate ; imprinting ; imprinting 00

Saccharomyces cerevisiaeSaccharomyces cerevisiae

In In Saccharomyces cerevisiaeSaccharomyces cerevisiae pheromones guide: pheromones guide:

- development of - development of mating projectionsmating projections

- intracellular migration of - intracellular migration of nucleusnucleus

Presence of N- and O-oligosaccharidesPresence of N- and O-oligosaccharides

-agglutinin-agglutinin (cell adhesion) – in the core region (cell adhesion) – in the core region 80-95% O-oligosach80-95% O-oligosach..

pheromones pheromones -agglutinin-gene-agglutinin-gene

cell-cell contactscell-cell contacts

Pheromones in lower levels of phylogeny (2)Pheromones in lower levels of phylogeny (2)

Pheromones in lower levels of phylogeny (3)Pheromones in lower levels of phylogeny (3)

Fungi – Fungi – Allomyces macrogynusAllomyces macrogynus

Sirenin Sirenin (1 pM – 1 (1 pM – 1 M)M) acts on gametsacts on gamets

HH++, K, K++, NH, NH44++, Na, Na++, Ca, Ca2+2+, Mg, Mg2+2+, La, La3+3+

- ions are - ions are repellent repellent itself itself - they can neutralize effects of each-other in competition- they can neutralize effects of each-other in competition

Practical applications of pheromonesPractical applications of pheromones

Artificial feritlization /inseminationArtificial feritlization /insemination

Anti-helminticAnti-helmintic

Insect repellentInsect repellent

Self – Non-self ?Self – Non-self ?

Male – Female ?Male – Female ?

„„T-shirt” testT-shirt” test

Pheromones in human (1)Pheromones in human (1)

Determination capacity of own-odor 75%Determination capacity of own-odor 75%

Distinguish male/female (female scores are better)Distinguish male/female (female scores are better)

Newborns – recognition of lactating breastNewborns – recognition of lactating breast

2 days2 days 002 weeks2 weeks + but no differentiation+ but no differentiation6 weeks6 weeks + mother preference+ mother preference

MatingMating

PheromonesPheromones

DogDogage (days)age (days) self self non-selfnon-self

20-2420-24 >>

31-36, 66-7231-36, 66-72 ~~

52-5652-56 malemale

Pheromone in hidePheromone in hide

The chair labelledThe chair labelled with male pheromonewith male pheromone was preferred by was preferred by female induvidualsfemale induviduals